N-hydroxylsulfonamide derivatives as new physiologically useful nitroxyl donors

ABSTRACT

The invention relates to N-hydroxylsulfonamide derivatives that donate nitroxyl (HNO) under physiological conditions and are useful in treating and/or preventing the onset and/or development of diseases or conditions that are responsive to nitroxyl therapy, including heart failure and ischemia/reperfusion injury. Novel N-hydroxylsulfonamide derivatives release NHO at a controlled rate under physiological conditions, and the rate of HNO release is modulated by varying the nature and location of functional groups on the N-hydroxylsulfonamide derivatives.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationSer. No. 60/995,636, filed Sep. 26, 2007 and entitled,“N-Hydroxylsulfonamide Derivatives as New Physiologically UsefulNitroxyl Donors,” which is incorporated herein by reference in itsentirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made in part with government support under Grant No.CHE-0518406 from the National Science Foundation. The government mayhave certain rights in this invention.

BACKGROUND OF THE INVENTION

Summary of Heart Failure

Congestive heart failure (CHF) is a generally progressive, lifethreatening condition in which myocardial contractility is depressedsuch that the heart is unable to adequately pump the blood returning toit, also referred to as decompensation. Symptoms include breathlessness,fatigue, weakness, leg swelling, and exercise intolerance. On physicalexamination, patients with heart failure often have elevated heart andrespiratory rates (an indication of fluid in the lungs), edema, jugularvenous distension, and enlarged hearts. The most common cause of CHF isatherosclerosis, which causes blockages in the coronary arteries thatprovide blood flow to the heart muscle. Ultimately, such blockages maycause myocardial infarction with subsequent decline in heart functionand resultant heart failure. Other causes of CHF include valvular heartdisease, hypertension, viral infections of the heart, alcoholconsumption, and diabetes. Some cases of CHF occur without clearetiology and are called idiopathic. The effects of CHF on a subjectexperiencing the condition can be fatal.

There are several types of CHF. Two types of CHF are identifiedaccording to which phase of the cardiac pumping cycle is more affected.Systolic heart failure occurs when the heart's ability to contractdecreases. The heart cannot pump with enough force to push a sufficientamount of blood into the circulation leading to a reduced leftventricular ejection fraction. Lung congestion is a typical symptom ofsystolic heart failure. Diastolic heart failure refers to the heart'sinability to relax between contractions and allow enough blood to enterthe ventricles. Higher filling pressures are required to maintaincardiac output, but contractility as measured by left ventricularejection fraction is typically normal. Swelling (edema) in the abdomenand legs is a typical symptom of diastolic heart failure. Often, anindividual experiencing heart failure will have some degree of bothsystolic heart failure and diastolic heart failure.

CHF is also classified according to its severity. The New York HeartAssociation classifies CHF into four classes: Class I involves noobvious symptoms, with no limitations on physical activity; Class IIinvolves some symptoms during or after normal activity, with mildphysical activity limitations; Class III involves symptoms with lessthan ordinary activity, with moderate to significant physical activitylimitations; and Class IV involves significant symptoms at rest, withsevere to total physical activity limitations. Typically, an individualprogresses through the classes as they live with the condition.

Although CHF is generally thought of as a chronic, progressivecondition, it can also develop suddenly. This type of CHF is calledacute CHF, and it is a medical emergency. Acute CHF can be caused byacute myocardial injury that affects either myocardial performance, suchas myocardial infarction, or valvular/chamber integrity, such as mitralregurgitation or ventricular septal rupture, which leads to an acuterise in left ventricular and diastolic pressure resulting in pulmonaryedema and dyspnea.

Common treatment agents for CHF include vasodilators (drugs that dilateblood vessels), positive inotropes (drugs that increase the heart'sability to contract), and diuretics (drugs to reduce fluid).Additionally, beta-antagonists (drugs that antagonize beta-adrenergicreceptors) have become standard agents for treating mild to moderateheart failure. Lowes et al, Clin. Cardiol., 23:III11-6 (2000).

Positive inotropic agents include beta-adrenergic agonists, such asdopamine, dobutamine, dopexamine, and isoproterenol. However, use of abeta-agonist has potential complications, such as arrhythmogenesis andincreased oxygen demand by the heart. Additionally, the initialshort-lived improvement of myocardial contractility afforded by thesedrugs is followed by an accelerated mortality rate resulting largelyfrom a greater frequency of sudden death. Katz, HEART FAILURE:PATHOPHYSIOLOGY, MOLECULAR BIOLOGY AND CLINICAL MANAGEMENT, Lippincott,Williams & Wilkins (1999).

Beta-antagonists antagonize beta-adrenergic receptor function. Whileinitially contra-indicated in heart failure, they have been found toprovide a marked reduction in mortality and morbidity in clinicaltrials. Bouzamondo et al., Fundam. Clin. Pharmacol., 15: 95-109 (2001).Accordingly, they have become an established therapy for heart failure.However, even subjects that improve under beta-antagonist therapy maysubsequently decompensate and require acute treatment with a positiveinotropic agent. Unfortunately, as their name suggests, beta-antagonistsblock the mechanism of action of the positive inotropic beta-agoniststhat are used in emergency care centers. Bristow et al., J. Card. Fail.,7: 8-12 (2001).

Vasodilators, such as nitroglycerin, have been used for a long period oftime to treat heart failure. However, the cause of nitroglycerin'stherapeutic effect was not known until late in the last century when itwas discovered that the nitric oxide molecule (NO) was responsible fornitroglycerin's beneficial effects. In some subjects experiencing heartfailure, a nitric oxide donor is administered in combination with apositive inotropic agent to both cause vasodilation and to increasemyocardial contractility. However, this combined administration canimpair the effectiveness of positive inotropic treatment agents. Forexample, Hart et al, Am. J. Physiol. Heart Circ. Physiol., 281:146-54(2001) reported that administration of the nitric oxide donor sodiumnitroprusside, in combination with the positive inotropic,beta-adrenergic agonist dobutamine, impaired the positive inotropiceffect of dobutamine. Hare et al., Circulation, 92:2198-203 (1995) alsodisclosed the inhibitory effect of nitric oxide on the effectiveness ofdobutamine.

As described in U.S. Pat. No. 6,936,639, compounds that donate nitroxyl(HNO) under physiological conditions have both positive inotropic andlusitropic effects and offer significant advantages over existingtreatments for failing hearts. Due to their concomitant positiveinotropic/lusotropic action and unloading effects, nitroxyl donors werereported as helpful in treating cardiovascular diseases characterized byhigh resistive load and poor contractile performance. In particular,nitroxyl-donating compounds were reported as useful in the treatment ofheart failure, including heart failure in individuals receivingbeta-antagonist therapy.

Summary of Ischemia

Ischemia is a condition characterized by an interruption or inadequatesupply of blood to tissue, which causes oxygen deprivation in theaffected tissue. Myocardial ischemia is a condition caused by a blockageor constriction of one or more of the coronary arteries, such as canoccur with atherosclerotic plaque occlusion or rupture. The blockade orconstriction causes oxygen deprivation of the non-perfused tissue, whichcan cause tissue damage. Further, upon reperfusion with subsequentreoxygenation of the tissue, when the blood is able to flow again or theoxygen demand of the tissue subsides, additional injury can be caused byoxidative stress.

Ischemia/reperfusion injury refers to tissue damage caused by oxygendeprivation followed by reoxygenation. The effects ofischemia/reperfusion injury in a subject experiencing the condition canbe fatal, particularly when the injury occurs in a critical organ suchas the heart or brain.

Accordingly, compounds and compositions effective in preventing orprotecting against ischemia/reperfusion injury would be usefulpharmaceuticals. Compounds such as nitroglycerin have been used for along period of time to help control vascular tone and protect againstmyocardial ischemia/reperfusion injury. It was discovered that thenitric oxide molecule was responsible for nitroglycerin's beneficialeffects. This discovery prompted interest in medical uses for nitricoxide and investigations into related species such as nitroxyl. Asreported in U.S. patent application Ser. No. 10/463,084 (U.S.Publication No. 2004/0038947) administration of a compound that donatesnitroxyl under physiological conditions, prior to ischemia, canattenuate ischemia/reperfusion injury to tissues, for example,myocardial tissues. This beneficial effect was reported as a surprisingresult given that nitroxyl was previously reported to increaseischemia/reperfusion injury (See, Ma et al., “Opposite Effects of NitricOxide and Nitroxyl on Postischemic Myocardial Injury,” Proc. Nat'l Acad.Sci., 96(25): 14617-14622 (1999), reporting that administration ofAngeli's salt (a nitroxyl donor under physiological conditions) toanesthetized rabbits during ischemia and 5 minutes prior to reperfusionincreased myocardial ischemia/reperfusion injury and Takahira et al.,“Dexamethasone Attenuates Neutrophil Infiltration in the Rat Kidney inIschemia/Reperfusion Injury: The Possible Role of Nitroxyl,” FreeRadical Biology & Medicine, 31(6):809-815 (2001) reporting thatadministration of Angeli's salt during ischemia and 5 minutes beforereperfusion of rat renal tissue contributed to neutrophil infiltrationinto the tissue, which is believed to mediate ischemia/reperfusioninjury). In particular, pre-ischemic administration of Angeli's salt andisopropylamine/NO has been reported to prevent or reduceischemia/reperfusion injury.

Summary of Nitroxyl Donors

To date, the vast majority of studies of the biological effect of HNOhave used the donor sodium dioxotrinitrate (“Angeli's salt” or “AS”).However, the chemical stability of AS has made it unsuitable to developas a therapeutic agent. N-hydroxybenzenesulfonamide (“Piloty's acid” or“PA”) has previously been shown to be a nitroxyl donor at high ph (>9)(Bonner, F. T.; Ko, Y. Inorg. Chem. 1992, 31, 2514-2519). However, underphysiological conditions, PA is a nitric oxide donor via an oxidativepathway (Zamora, R.; Grzesiok, A.; Weber, H.; Feelisch, M. Biochem. J.1995, 312, 333-339). Thus, the physiological effects of AS and PA arenot the same because AS is a nitroxyl donor under physiologicalconditions whereas PA is a nitric oxide donor under physiologicalconditions.

Although U.S. Pat. No. 6,936,639 and U.S. Publication No. 2004/0038947describe PA as a compound that donates nitroxyl and note that othersulfohydroxamic acids and their derivatives are therefore also useful asnitroxyl donors, PA does not in fact donate significant amounts ofnitroxyl under physiological conditions (See Zamora, supra).

Several substituted N-hydroxylbenzenesulfonamides have been reported asinhibitors of carbonic anhydrase, with no mention of HNO production(see, (a) Mincione, F.; Menabuoni, L.; Briganti., F; Mincione, G.;Scozzafava, A.; Supuran, C. T. J. Enzyme Inhibition 1998, 13, 267-284and (b) Scozzafava, A.; Supuran, C. T., J. Med. Chem. 2000, 43,3677-3687).

N-hydroxylsulfonamide derivatives as new physiologically useful nitroxyldonors are also described in PCT application No. PCT/US2007/006710 filedMar. 16, 2007. However, compounds of the formula (I) described thereinare not substituted with at least one carboxyl, carboxyl ester,acylamino or sulfonylamino group and compounds of the formula (II) or(III) described therein are not substituted with at least onecarbonylamino or sulfonylamino group.

Significant Medical Need

Despite efforts towards the development of new therapies for thetreatment of diseases and conditions such as heart failure andischemia/reperfusion injury, there remains a significant interest in andneed for additional or alternative compounds that treat or prevent theonset or severity of these and related diseases or conditions. Inparticular, there remains a significant medical need for alternative oradditional therapies for the treatment of diseases or conditions thatare responsive to nitroxyl therapy. New compounds that donate nitroxylunder physiological conditions and methods of using compounds thatdonate nitroxyl under physiological conditions may thus find use astherapies for treating, preventing and/or delaying the onset and/ordevelopment of diseases or conditions responsive to nitroxyl therapy,including heart disease and ischemia/reperfusion injury. Preferably, thetherapeutic agents can improve the quality of life and/or prolong thesurvival time for patients with the disease or condition.

BRIEF SUMMARY OF THE INVENTION

Methods, compounds and compositions for treating and/or preventing theonset or development of diseases or conditions that are responsive tonitroxyl therapy are described. Aromatic and non-aromaticN-hydroxylsulfonamide derivatives that donate nitroxyl underphysiological conditions are described. By modifying PA with appropriatesubstituents, such as electron-withdrawing groups or groups thatsterically hinder the sulfonyl moiety, the HNO producing capacity ofthese derivatives is substantially enhanced under physiologicalconditions. Significantly, when compared to AS, PA has the capacity forbroad substituent modification, enabling optimization of physicochemicaland pharmacological properties. Such optimization is reported herein.

In one embodiment, the present invention provides a method ofadministering to a subject in need thereof, a therapeutically effectiveamount of a derivative of PA wherein the derivative donates nitroxylunder physiological conditions. In one embodiment, the inventionembraces a method of treating or preventing the onset and/or developmentof a disease or condition that is responsive to nitroxyl therapy, themethod comprising administering to an individual in need thereof anN-hydroxylsulfonamide that donates an effective amount of nitroxyl underphysiological conditions. Also embraced are methods of treating heartfailure or ischemia/reperfusion injury by administering to an individualin need thereof an N-hydroxysulfonamide that donates an effective amountof nitroxyl under physiological conditions.

Kits comprising the compounds are also described, which may optionallycontain a second therapeutic agent such as a positive inotropiccompound, which may be, e.g., a beta-adrenergic receptor agonist.

Novel compounds that find use in the invention described herein includecompounds of the formula (I), (II) or (III):

where R¹ is H; R² is H, aralkyl or heterocyclyl; m and n areindependently an integer from 0 to 2; x is an integer from 0 to 4 and yis an integer from 0 to 3, provided that at least one of x and y isgreater than 0; b is an integer from 1-4; R³, R⁴, R⁵, R⁶ and R⁷ areindependently selected from the group consisting of H, halo,alkylsulfonyl, N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl, cyano,alkoxy, perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl,alkylsulfinyl, heterocycloalkyl, substituted heterocycloalkyl,dialkylamino, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carboxyl, carboxyl ester, acylamino and sulfonylamino,provided that at least one of R³, R⁴, R⁵, R⁶ and R⁷ is carboxyl,carboxyl ester, acylamino or sulfonylamino; each R⁸ and R⁹ isindependently selected from the group consisting of halo, alkylsulfonyl,N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl, cyano, alkoxy,perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl, alkylsulfinyl,heterocycloalkyl, substituted heterocycloalkyl, dialkylamino, NH₂, OH,C(O)OH, C(O)Oalkyl, NHC(O)alkylC(O)OH, C(O)NH₂, NHC(O)alkylC(O)alkyl,NHC(O)alkenylC(O)OH, NHC(O)NH₂, OalkylC(O)Oalkyl, NHC(O)alkyl,C(═N—OH)NH₂, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carbonylamino, and sulfonylamino, provided that: (1) atleast one R⁸ is carbonylamino or sulfonylamino when the compound is ofthe formula (III) and (2) at least one of R⁸ and R⁹ is carbonylamino orsulfonylamino when the compound is of the formula (II); A is acycloalkyl, heterocycloalkyl, aromatic or heteroaromatic ring containingring moieties Q¹, Q², Q³ and Q⁴, which are taken together with V and Wto form ring A; B is a cycloalkyl, heterocycloalkyl, aromatic orheteroaromatic ring containing ring moieties Q⁵, Q⁶, Q⁷ and Q⁸, whichare taken together with the V and W to form ring B; V and W areindependently C, CH, N or NR¹⁰; Q¹, Q², Q³, Q⁴, Q⁵, Q⁶, Q⁷ and Q⁸ areindependently selected from the group consisting of C, CH₂, CH, N, NR¹⁰,O and S; C is a heteroaromatic ring containing ring moieties Q⁹, Q¹⁰,Q¹¹, Q¹², Q¹³ and Q¹⁴ that are independently selected from the groupconsisting of C, CH₂, CH, N, NR¹⁰, O and S, provided that at least oneof Q⁹, Q¹⁰, Q¹¹, Q¹², Q¹³ and Q¹⁴ is N, NR¹⁰, O or S; and R¹⁰ is H,alkyl, acyl or sulfonyl. Pharmaceutically acceptable salts of any of theforegoing are also described.

In one variation, the compound is of the formula (I), (II) or (III)where R¹ is H; R² is H; m and n are independently an integer from 0 to2; x is an integer from 0 to 4 and y is an integer from 0 to 3, providedthat at least one of x and y is greater than 0; b is an integer from1-4; R³, R⁴, R⁵, R⁶ and R⁷ are independently selected from the groupconsisting of H, halo, alkylsulfonyl, substituted alkylsulfonyl,N-hydroxylsulfonamidyl, substituted N-hydroxylsulfonamidyl,perhaloalkyl, substituted perhaloalkyl (where one or more halo may besubstituted with a substituent), nitro, aryl, substituted aryl, cyano,alkoxy, substituted alkoxy, perhaloalkoxy, substituted perhaloalkoxy,alkyl, substituted alkyl, aryloxy, substituted aryloxy, alkylsulfanyl,substituted alkylsulfanyl, alkylsulfinyl, substituted alkylsulfinyl,heterocycloalkyl, substituted heterocycloalkyl, dialkylamino,substituted dialkylamino, cycloalkoxy, substituted cycloalkoxy,cycloalkylsulfanyl, substituted cycloalkylsulfanyl, arylsulfanyl,substituted arylsulfanyl, arylsulfinyl, substituted arylsulfinyl,carboxyl, carboxyl ester, acylamino and sulfonylamino, provided that atleast one of R³, R⁴, R⁵, R⁶ and R⁷ is carboxyl, carboxyl ester,acylamino or sulfonylamino; each R⁸ and R⁹ is independently selectedfrom the group consisting of halo, alkylsulfonyl, substitutedalkylsulfonyl, N-hydroxylsulfonamidyl, substitutedN-hydroxylsulfonamidyl, perhaloalkyl, substituted perhaloalkyl, nitro,aryl, substituted aryl, cyano, alkoxy, substituted alkoxy,perhaloalkoxy, substituted perhaloalkoxy, alkyl, substituted alkyl,aryloxy, substituted aryloxy, alkylsulfanyl, substituted alkylsulfanyl,alkylsulfinyl, substituted alkylsulfinyl, heterocycloalkyl, substitutedheterocycloalkyl, dialkylamino, substituted dialkylamino, NH₂, OH,C(O)OH, C(O)Oalkyl, NHC(O)alkylC(O)OH, C(O)NH₂, NHC(O)alkylC(O)alkyl,NHC(O)alkenylC(O)OH, NHC(O)NH₂, OalkylC(O)Oalkyl, NHC(O)alkyl,C(═N—OH)NH₂, cycloalkoxy, substituted cycloalkoxy, cycloalkylsulfanyl,substituted cycloalkylsulfanyl, arylsulfanyl, substituted arylsulfanyl,arylsulfinyl, substituted arylsulfinyl (where any listing of alkyl oralkenyl in the moieties above intends unsubstituted or substituted alkylor alkenyl), carbonylamino and sulfonylamino, provided that: (1) atleast one R⁸ is carbonylamino or sulfonylamino when the compound is ofthe formula (III) and (2) at least one of R⁸ and R⁹ is carbonylamino orsulfonylamino when the compound is of the formula (II); A is acycloalkyl, heterocycloalkyl, aromatic or heteroaromatic ring containingring moieties Q¹, Q², Q³ and Q⁴, which are taken together with V and Wto form ring A; B is a cycloalkyl, heterocycloalkyl, aromatic orheteroaromatic ring containing ring moieties Q⁵, Q⁶, Q⁷ and Q⁸, whichare taken together with the V and W to form ring B; V and W areindependently C, CH, N or NR¹⁰; Q¹, Q², Q³, Q⁴, Q⁵, Q⁶, Q⁷ and Q⁸ areindependently selected from the group consisting of C, CH₂, CH, N, NR¹⁰,O and S; C is a heteroaromatic ring containing ring moieties Q⁹, Q¹⁰,Q¹¹, Q¹², Q¹³ and Q¹⁴ that are independently selected from the groupconsisting of C, CH₂, CH, N, NR¹⁰, O and S, provided that at least oneof Q⁹, Q¹⁰, Q¹¹, Q¹², Q¹³ and Q¹⁴ is N, NR¹⁰, O or S; and R¹⁰ is H,alkyl, acyl or sulfonyl. Pharmaceutically acceptable salts of any of theforegoing are also described.

Methods of using the compounds detailed herein are also described,including a method of treating, preventing or delaying the onset ordevelopment of a disease or condition that is responsive to nitroxyltherapy, comprising administering to an individual in need thereof acompound of the invention that donates nitroxyl under physiologicalconditions or a pharmaceutically acceptable salt thereof.

Pharmaceutical compositions comprising a compound of the invention aredisclosed, such as pharmaceutical compositions that are amenable tointravenous injection. Kits comprising a compound of the invention andinstructions for use are also described.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the nitrous oxide evolved from Compounds 1-5 as apercent of Angeli's Salt and the mole percent of N₂O generated per moleof sample for compounds 1-5.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless clearly indicated otherwise, the following terms as used hereinhave the meanings indicated below.

Use of the terms “a”, “an” and the like refers to one or more.

“Aralkyl” refers to a residue in which an aryl moiety is attached to theparent structure via an alkyl residue. Examples include benzyl(—CH₂-Ph), phenethyl (—CH₂CH₂Ph), phenylvinyl (—CH═CH-Ph), phenylallyland the like.

“Acyl” refers to and includes the groups —C(O)H, —C(O)alkyl,—C(O)substituted alkyl, —C(O)alkenyl, —C(O)substituted alkenyl,—C(O)alkynyl, —C(O)substituted alkynyl, —C(O)cycloalkyl,—C(O)substituted cycloalkyl, —C(O)aryl, —C(O)substituted aryl,—C(O)heteroaryl, —C(O)substituted heteroaryl, —C(O)heterocyclic, and—C(O)substituted heterocyclic wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein or otherwise known in the art.

“Heterocyclyl” or “Heterocycloalkyl” refers to a cycloalkyl residue inwhich one to four of the carbons is replaced by a heteroatom such asoxygen, nitrogen or sulfur. Examples of heterocycles whose radicals areheterocyclyl groups include tetrahydropyran, morpholine, pyrrolidine,piperidine, thiazolidine, oxazole, oxazoline, isoxazole, dioxane,tetrahydrofuran and the like. A specific example of a heterocyclylresidue is tetrahydropyran-2-yl.

“Substituted heterocyclo” or “substituted heterocylcoalkyl” refers to anheterocyclyl group having from 1 to 5 substituents. For instance, aheterocyclyl group substituted with 1 to 5 groups such as halo, nitro,cyano, oxo, aryl, alkoxy, alkyl, acyl, acylamino, amino, hydroxyl,carboxyl, carboxylalkyl, thiol, thioalkyl, heterocyclyl, —OS(O)₂-alkyl,and the like is a substituted alkyl. A particular example of asubstituted heterocylcoalkyl is N-methylpiperazino.

“Alkyl” intends linear hydrocarbon structures having 1 to 20 carbonatoms, preferably 1 to 12 carbon atoms and more preferably 1 to 8 carbonatoms. Alkyl groups of fewer carbon atoms are embraced, such asso-called “lower alkyl” groups having 1 to 4 carbon atoms. “Alkyl” alsointends branched or cyclic hydrocarbon structures having 3 to 20 carbonatoms, preferably 3 to 12 carbon atoms and more preferably 3 to 8 carbonatoms. For any use of the term “alkyl,” unless clearly indicatedotherwise, it is intended to embrace all variations of alkyl groupsdisclosed herein, as measured by the number of carbon atoms, the same asif each and every alkyl group was explicitly and individually listed foreach usage of the term. For instance, when a group such as R³ may be an“alkyl,” intended is a C₁-C₂₀ alkyl or a C₁-C₁₂ alkyl or a C₁-C₈ alkylor a lower alkyl or a C₂-C₂₀ alkyl or a C₃-C₁₂ alkyl or a C₃-C₈ alkyl.The same is true for other groups listed herein, which may includegroups under other definitions, where a certain number of atoms islisted in the definition. When the alkyl group is cyclic, it may also bereferred to as a cycloalkyl group and have e.g., 3 to 20 annular carbonatoms, preferably 3 to 12 annular carbon atoms and more preferably 3 to8 annular carbon atoms. When an alkyl residue having a specific numberof carbons is named, all geometric isomers having that number of carbonsare intended to be encompassed; thus, for example, “butyl” is meant toinclude n-butyl, sec-butyl, iso-butyl and t-butyl; “propyl” includesn-propyl and iso-propyl. Examples of alkyl groups include methyl, ethyl,n-propyl, i-propyl, t-butyl, n-heptyl, octyl, cyclopentyl, cyclopropyl,cyclobutyl, norbornyl, and the like. One or more degrees of unsaturationmay occur in an alkyl group. Thus, an alkyl group also embraces alkenyland alkynyl residues. “Alkenyl” is understood to refer to a group of 2or more carbon atoms, such as 2 to 10 carbon atoms and more preferably 2to 6 carbon atoms and having at least 1 and preferably from 1-2 sites ofalkenyl unsaturation. Examples of an alkenyl group include —C═CH₂,—CH₂CH═CHCH₃ and —CH₂CH═CH—CH═CH₂. “Alkynyl” refers to alkynyl grouppreferably having from 2 to 10 carbon atoms and more preferably 3 to 6carbon atoms and having at least 1 and preferably from 1-2 sites ofalkynyl unsaturation, such as the moiety —C≡CH. Alkyl is also usedherein to denote an alkyl residue as part of a larger functional groupand when so used, is taken together with other atoms to form anotherfunctional group. For instance, reference to —C(O)Oalkyl intends anester functional group, where the alkyl portion of the moiety may be anyalkyl group, and provide by way of example only, the functional group—C(O)OCH₃, —C(O)(O)CH═CH₂ and the like. Another example of an alkylgroup as part of a larger structure includes the residue—NHC(O)alkylC(O)OH, which e.g., may be NHC(O)CH₂CH₂C(O)OH when alkyl is—CH₂CH₂—.

“Substituted alkyl” refers to an alkyl group having from 1 to 5substituents. For instance, an alkyl group substituted with a group suchas halo, nitro, cyano, oxo, aryl, alkoxy, acyl, acylamino, amino,hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkyl, heterocyclyl,—OS(O)₂-alkyl, and the like is a substituted alkyl. Likewise,“substituted alkenyl” and “substituted alkynyl” refer to alkenyl oralkynyl groups having 1 to 5 substituents.

As used herein the term “substituent” or “substituted” means that ahydrogen radical on a compound or group (such as, for example, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl,heteroaryl, substituted heteroaryl, heteroaralkyl, substitutedheteroaralkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl,substituted heterocycloalkyl, heterocyclyl and substituted heterocyclyl)is replaced with any desired group that does not substantially adverselyaffect the stability of the compound. In one embodiment, desiredsubstituents are those which do not adversely affect the activity of acompound. The term “substituted” refers to one or more substituents(which may be the same or different), each replacing a hydrogen atom.Examples of substituents include, but are not limited to, halogen (F,Cl, Br, or I), hydroxyl, amino, alkylamino, arylamino, dialkylamino,diarylamino, cyano, nitro, mercapto, oxo (i.e., carbonyl), thio, imino,formyl, carbamido, carbamyl, carboxyl, thioureido, thiocyanato,sulfoamido, sulfonylalkyl, sulfonylaryl, alkyl, alkenyl, alkoxy,mercaptoalkoxy, aryl, heteroaryl, cyclyl, heterocyclyl, wherein alkyl,alkenyl, alkyloxy, aryl, heteroaryl, cyclyl, and heterocyclyl areoptionally substituted with alkyl, aryl, heteroaryl, halogen, hydroxyl,amino, mercapto, cyano, nitro, oxo (═O), thioxo (═S), or imino(═Nalkyl). In other embodiments, substituents on any group (such as, forexample, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, aryl, substituted aryl, aralkyl,substituted aralkyl, heteroaryl, substituted heteroaryl, heteroaralkyl,substituted heteroaralkyl, cycloalkyl, substituted cycloalkyl,heterocycloalkyl, substituted heterocycloalkyl, heterocyclyl andsubstituted heterocyclyl) can be at any atom of that group (such as on acarbon atom of the primary carbon chain of a substituted alkyl group oron a substituent already present on a substituted alkyl group) or at anyatom of, wherein any group that can be substituted (such as, forexample, alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl,heteroaralkyl, cycloalkyl, cyclyl, heterocycloalkyl, and heterocyclyl)can be optionally substituted with one or more substituents (which maybe the same or different), each replacing a hydrogen atom. Examples ofsuitable substituents include, but not limited to alkyl, alkenyl,alkynyl, cyclyl, cycloalkyl, heterocyclyl, heterocycloalkyl, aralkyl,heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro,alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl),carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl,alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl,thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl,dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, oralkoxycarbonylamino; alkylamino, arylamino, diarylamino, alkylcarbonyl,or arylamino-substituted aryl; arylalkylamino, aralkylaminocarbonyl,amido, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl,alkylsulfonylamino, arylsulfonylamino, imino, carbamido, carbamyl,thioureido, thiocyanato, sulfoamido, sulfonylalkyl, sulfonylaryl, ormercaptoalkoxy. Additional suitable substituents on alkyl, alkenyl,alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, cyclyl,heterocycloalkyl, and heterocyclyl include, without limitation halogen,CN, NO₂, OR¹¹, SR¹¹, S(O)₂OR¹¹, NR¹¹R¹², C₁-C₂ perfluoroalkyl, C₁-C₂perfluoroalkoxy, 1,2-methylenedioxy, (═O), (═S), (═NR¹¹), C(O)OR¹¹,C(O)R¹¹R¹², OC(O)NR¹¹R¹², NR¹¹C(O)NR¹¹R¹², C(NR¹²)NR¹¹R¹²,NR¹¹C(NR¹²)NR¹¹R¹², S(O)₂NR¹¹R¹²R¹³, C(O)H, C(O)R¹³, NR¹¹C(O)R¹³,Si(R¹¹)₃, OSi(R¹¹)₃, Si(OH)₂R¹¹, B(OH)₂, P(O)(OR¹¹)₂, S(O)R¹³, orS(O)₂R¹³. Each R¹¹ is independently hydrogen, C₁-C₆ alkyl optionallysubstituted with cycloalkyl, aryl, heterocyclyl, or heteroaryl. Each R¹²is independently hydrogen, C₃-C₆ cycloalkyl, aryl, heterocyclyl,heteroaryl, C₁-C₄ alkyl or C₁-C₄ alkyl substituted with C₃-C₆cycloalkyl, aryl, heterocyclyl or heteroaryl. Each R¹³ is independentlyC₃-C₆ cycloalkyl, aryl, heterocyclyl, heteroaryl, C₁-C₄ alkyl or C₁-C₄alkyl substituted with C₃-C₆ cycloalkyl, aryl, heterocyclyl orheteroaryl. Each C₃-C₆ cycloalkyl, aryl, heterocyclyl, heteroaryl andC₁-C₄ alkyl in each R¹¹, R¹² and R¹³ can optionally be substituted withhalogen, CN, C₁-C₄ alkyl, OH, C₁-C₄ alkoxy, COOH, C(O)OC₁-C₄ alkyl, NH₂,C₁-C₄ alkylamino, or C₁-C₄ dialkylamino. Substituents can also be“electron-withdrawing groups.”

“Electron withdrawing group” refers to groups that reduce electrondensity of the moiety to which they are attached (relative to thedensity of the moiety without the substituent). Such groups include, forexample, F, Cl, Br, I, —CN, —CF₃, —NO₂, —SH, —C(O)H, —C(O)alkyl,—C(O)Oalkyl, —C(O)OH, —C(O)Cl, —S(O)₂OH, —S(O)₂NHOH, —NH₃ and the like.

“Halo” refers to fluorine, chlorine, bromine or iodine.

“Alkylsulfonyl” refers to groups —SO₂alkyl and —SO₂substituted alkyl,which includes the residues —SO₂cycloalkyl, —SO₂substituted cycloalkyl,—SO₂alkenyl, —SO₂substituted alkenyl, —SO₂alkynyl, —SO₂substitutedalkynyl, where alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl and substituted cycloalkyl areas defined herein.

“N-hydroxylsulfonamidyl” refers to —S(O)₂NROH, where R is H or alkyl.

“Perhaloalkyl” refers to an alkyl group where each H of the hydrocarbonis replaced with F. Examples of perhalo groups include —CF₃ and —CF₂CF₃.

“Aryl” intends a monocyclic, bicyclic or tricyclic aromatic ring. Anaryl group is preferably a 5- or 6-membered aromatic or heteroaromaticring containing 0-3 annular heteroatoms selected from O, N, or S; abicyclic 9- or 10-membered aromatic or heteroaromatic ring system(meaning the ring system has 9 or 10 annular atoms) containing 0-3annular heteroatoms selected from O, N, or S; or a tricyclic 13- or14-membered aromatic or heteroaromatic ring system (meaning the ringsystem has 13 or 14 annular atoms) containing 0-3 annular heteroatomsselected from O, N, or S. Examples of groups whose radicals are arylgroups include e.g., benzene, naphthalene, indane, tetralin, imidazole,pyridine, indole, thiophene, benzopyranone, thiazole, furan,benzimidazole, benzoxazole, benzthiazole, quinoline, isoquinoline,quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.

“Substituted aryl” refers to a group having from 1 to 3 substituents.For instance, an aryl group substituted with 1 to 3 groups such as halo,nitro, cyano, oxo, aryl, alkoxy, alkyl, acyl, acylamino, amino,hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkyl, heterocyclyl,—OS(O)₂-alkyl, and the like is a substituted aryl.

“Alkoxy” refers to an alkyl group that is connected to the parentstructure through an oxygen atom (—O-alkyl). When a cycloalkyl group isconnected to the parent structure through an oxygen atom, the group mayalso be referred to as a cycloalkoxy group. Examples include methoxy,ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy and the like.A “perhaloalkoxy” intends a perhaloalkyl group attached to the parentstructure through an oxygen, such as the residue —O—CF₃.

“Aryloxy” refers to an aryl group that is connected to the parentstructure through an oxygen atom (—O-aryl), which by way of exampleincludes the residues phenoxy, naphthoxy, and the like. “Substitutedaryloxy” refers to a substituted aryl group connected to the parentstructure through an oxygen atom (—O-substituted aryl).

“Alkylsulfanyl” refers to an alkyl group that is connected to the parentstructure through a sulfur atom (—S-alkyl) and refers to groups —S-alkyland —S-substituted alkyl, which includes the residues —S-cycloalkyl,—S-substituted cycloalkyl, —S-alkenyl, —S-substituted alkenyl,—S-alkynyl, —S-substituted alkynyl, where alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyland substituted cycloalkyl are as defined herein. When a cycloalkylgroup is connected to the parent structure through an sulfur atom, thegroup may also be referred to as a cycloalkylsulfanyl group. By way ofexample, alkylsulfanyl includes —S—CH(CH₃), —S—CH₂CH₃ and the like.

“Alkylsulfinyl” refers to an alkyl group that is connected to the parentstructure through a S(O) moiety and refers to groups —S(O)alkyl and—S(O)substituted alkyl, which includes the residues —S(O)cycloalkyl,—S(O)substituted cycloalkyl, —S(O)alkenyl, —S(O)substituted alkenyl,—S(O)alkynyl, —S(O)substituted alkynyl, where alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyland substituted cycloalkyl are as defined herein. By way of example,alkylsulfinyl includes the residues —S(O)CH(CH₃), —S(O)CH₃,—S(O)cyclopentane and the like.

“Arylsulfinyl” refers to an aryl group that is connected to the parentstructure through a S(O) moiety, which by way of example includes theresidue —S(O)Ph.

“Dialkylamino” refers to the group —NR₂ where each R is an alkyl group.Examples of dialkylamino groups include —N(CH₃)₂, —N(CH₂CH₂CH₂CH₃)₂, andN(CH₃)(CH₂CH₂CH₂CH₃).

“Carboxyl” refers to —C(O)OH.

“Carboxyl ester” as used herein refers to the groups —C(O)O-alkyl,—C(O)O-substituted alkyl, —C(O)O-aryl, —C(O)O-substituted aryl,—C(O)O-alkenyl, —C(O)O-substituted alkenyl, —C(O)O-alkynyl,—C(O)O-substituted alkynyl, —C(O)O-heteroaryl, —C(O)O-substitutedheteroaryl, —C(O)O-heterocyclic or —C(O)O-substituted heterocyclic.

“Acylamino” refers to the group —C(O)NR_(a)R_(b) where each R_(a) andR_(b) group is independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic or R_(a)and R_(b) groups can be joined together with the nitrogen atom to form aheterocyclic or substituted heterocyclic ring. An examples of anacylamino moiety includes —C(O)morpholino.

“Sulfonylamino” refers to the groups —SO₂NH₂, —SO₂NR-alkyl,—SO₂NR-substituted alkyl, —SO₂NR-alkenyl, —SO₂NR-substituted alkenyl,—SO₂NR-alkynyl, —SO₂NR-substituted alkynyl, —SO₂NR-aryl,—SO₂NR-substituted aryl, —SO₂NR-heteroaryl, —SO₂NR-substitutedheteroaryl, —SO₂NR-heterocyclic, and —SO₂NR-substituted heterocyclicwhere R is hydrogen or alkyl, or —SO₂NR₂, where the two R groups aretaken together and with the nitrogen atom to which they are attached toform a heterocyclic or substituted heterocyclic ring.

“Carbonylamino” refers to the groups —CONH₂, —CONR-alkyl,—CONR-substituted alkyl, —CONR-alkenyl, —CONR-substituted alkenyl,—CONR-alkynyl, —CONR-substituted alkynyl, —CONR-aryl, —CONR-substitutedaryl, —CONR-heteroaryl, —CONR-substituted heteroaryl,—CONR-heterocyclic, and —CONR-substituted heterocyclic where R ishydrogen or alkyl, or —CONR₂, where the two R groups are taken togetherand with the nitrogen atom to which they are attached to form aheterocyclic or substituted heterocyclic ring.

“Pharmaceutically acceptable salt” refers to pharmaceutically acceptablesalts of a compound described herein, such as a compound of Formula (I),(II) or (III) or other nitroxyl donor of the invention, which salts maybe derived from a variety of organic and inorganic counter ions wellknown in the art and include, by way of example only, sodium, potassium,calcium, magnesium, ammonium, tetraalkylammonium, and the like; when themolecule contains a basic functionality, salts of organic or inorganicacids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate,maleate, oxalate and the like. Illustrative salts include, but are notlimited, to sulfate, citrate, acetate, chloride, bromide, iodide,nitrate, bisulfate, phosphate, acid phosphate, lactate, salicylate, acidcitrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate,succinate, maleate, besylate, fumarate, gluconate, glucaronate,saccharate, formate, benzoate, glutamate, methanesulfonate,ethanesulfonate, benzenesulfonate, and p-toluenesulfonate salts.Accordingly, a salt may be prepared from a compound of any one of theformulae disclosed herein having an acidic functional group, such as acarboxylic acid functional group, and a pharmaceutically acceptableinorganic or organic base. Suitable bases include, but are not limitedto, hydroxides of alkali metals such as sodium, potassium, and lithium;hydroxides of alkaline earth metal such as calcium and magnesium;hydroxides of other metals, such as aluminum and zinc; ammonia, andorganic amines, such as unsubstituted or hydroxy-substituted mono-, di-,or trialkylamines; dicyclohexylamine; tributyl amine; pyridine;N-methyl, N-ethylamine; diethylamine; triethylamine; mono-, bis-, ortris-(2-hydroxy-lower alkyl amines), such as mono-, bis-, ortris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, ortris-(hydroxymethyl)methylamine, N,N,-di-lower alkyl-N-(hydroxy loweralkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl) amine, ortri-(2-hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such asarginine, lysine, and the like. A salt may also be prepared from acompound of any one of the formulae disclosed herein having a basicfunctional group, such as an amino functional group, and apharmaceutically acceptable inorganic or organic acid. Suitable acidsinclude hydrogen sulfate, citric acid, acetic acid, hydrochloric acid(HCl), hydrogen bromide (HBr), hydrogen iodide (HI), nitric acid,phosphoric acid, lactic acid, salicylic acid, tartaric acid, ascorbicacid, succinic acid, maleic acid, besylic acid, fumaric acid, gluconicacid, glucaronic acid, formic acid, benzoic acid, glutamic acid,methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, andp-toluenesulfonic acid.

Unless clearly indicated otherwise, “an individual” as used hereinintends a mammal, including but not limited to a human.

The term “effective amount” intends such amount of a compound or apharmaceutically acceptable salt thereof, which in combination with itsparameters of efficacy and toxicity, as well as based on the knowledgeof the practicing specialist should be effective in a given therapeuticform. As is understood in the art, an effective amount may be in one ormore doses.

As used herein, “treatment” or “treating” is an approach for obtaining abeneficial or desired result, including clinical results. For purposesof this invention, beneficial or desired results include but are notlimited to inhibiting and/or suppressing the onset and/or development ofa disease or condition that is responsive to nitroxyl therapy orreducing the severity of such disease or condition, such as reducing thenumber and/or severity of symptoms associated with the disease orcondition, increasing the quality of life of those suffering from thedisease or condition, decreasing the dose of other medications requiredto treat the disease or condition, enhancing the effect of anothermedication an individual is taking for the disease or condition andprolonging survival of individuals having the disease or condition. Thedisease or condition may be a cardiovascular disease or condition, whichincludes, but is not limited to, coronary obstructions, coronary arterydisease (CAD), angina, heart attack, myocardial infarction, high bloodpressure, ischemic cardiomyopathy and infarction, diastolic heartfailure, pulmonary congestion, pulmonary edema, cardiac fibrosis,valvular heart disease, pericardial disease, circulatory congestivestates, peripheral edema, ascites, Chagas' disease, ventricularhypertrophy, heart valve disease, heart failure, including but notlimited to congestive heart failure such as acute congestive heartfailure and acute decompensated heart failure. Related symptoms that maybe alleviated by the methods herein include shortness of breath,fatigue, swollen ankles or legs, angina, loss of appetite, weight gainor loss, associated with aforementioned diseases or disorders. Thedisease or condition may involve ischemia/reperfusion injury.

As used herein, “preventing” refers to reducing the probability ofdeveloping a disorder or condition in an individual who does not have,but is at risk of developing a disorder or condition.”

An individual “at risk” may or may not have a detectable disease orcondition, and may or may not have displayed a detectable disease orcondition prior to the treatment methods described herein. “At risk”denotes that an individual has one or more so-called risk factors, whichare measurable parameters that correlate with development of a diseaseor condition and are known in the art. An individual having one or moreof these risk factors has a higher probability of developing the diseaseor condition than an individual without these risk factor(s).

“Nitroxyl” refers to the species HNO.

As used herein, a compound is a “nitroxyl donor” if it donates nitroxylunder physiological conditions. As used herein, nitroxyl donors of theinvention may alternatively be referred to as “a compound” or “thecompound.” Preferably, the nitroxyl donor is capable of donating aneffective amount of nitroxyl in vivo and has a safety profile indicatingthe compound would be tolerated by an individual in the amount necessaryto achieve a therapeutic effect. One of ordinary skill in the art wouldbe able to determine the safety of administering particular compoundsand dosages to live subjects. One of skill in the art may also determinewhether a compound is a nitroxyl donor by evaluating whether it releasesHNO under physiological conditions. Compounds are easily tested fornitroxyl donation with routine experiments. Although it is impracticalto directly measure whether nitroxyl is donated, several tests areaccepted for determining whether a compound donates nitroxyl. Forexample, the compound of interest can be placed in solution, for examplein water, in a sealed container. After sufficient time fordisassociation has elapsed, such as from several minutes to severalhours, the headspace gas is withdrawn and analyzed to determine itscomposition, such as by gas chromatography and/or mass spectroscopy. Ifthe gas N₂O is formed (which occurs by HNO dimerization), the test ispositive for nitroxyl donation and the compound is a nitroxyl donor. Thelevel of nitroxyl donating ability may be expressed as a percentage of acompound's theoretical maximum. A compound that donates a “significantlevel of nitroxyl” intends a compound that donates 40% or more or 50% ormore of its theoretical maximum amount of nitroxyl. In one variation,the compounds for use herein donate 60% or more of the theoreticalmaximum amount of nitroxyl. In another variation, the compounds for useherein donate 70% or more of the theoretical maximum amount of nitroxyl.In another variation, the compounds for use herein donate 80% or more ofthe theoretical maximum amount of nitroxyl. In another variation, thecompounds for use herein donate 90% or more of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 70% and about 90% of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 85% and about 95% of the theoretical maximumamount of nitroxyl. In yet another variation, the compounds for useherein donate between about 90% and about 95% of the theoretical maximumamount of nitroxyl. Compounds that donate less than 40% or less than 50%of their theoretical amount of nitroxyl are still nitroxyl donors andmay be used in the invention disclosed herein. A compound that donatesless than 50% of the theoretical amount of nitroxyl may be used in themethods described, and may require higher dosing levels as compared tocompounds that donate a significant level of nitroxyl. Nitroxyl donationalso can be detected by exposing the test compound to metmyoglobin(Mb³⁺). Nitroxyl reacts with Mb³⁺ to form an Mb²⁺-NO complex, which canbe detected by changes in the ultraviolet/visible spectrum or byElectron Paramagnetic Resonance (EPR). The Mb²⁺-NO complex has an EPRsignal centered around a g-value of about 2. Nitric oxide, on the otherhand, reacts with Mb³⁺ to form an Mb³⁺-NO complex that is EPR silent.Accordingly, if the candidate compound reacts with Mb³⁺ to form acomplex detectable by common methods such as ultraviolet/visible or EPR,then the test is positive for nitroxyl donation. Testing for nitroxyldonation may be performed at physiologically relevant pH.

A “positive inotrope” as used herein is an agent that causes an increasein myocardial contractile function. Such an agent includes abeta-adrenergic receptor agonist, an inhibitor of phosphodiesteraseactivity, and calcium-sensitizers. Beta-adrenergic receptor agonistsinclude, among others, dopamine, dobutamine, terbutaline, andisoproterenol. Analogs and derivatives of such compounds are alsointended. For example, U.S. Pat. No. 4,663,351 describes a dobutamineprodrug that can be administered orally. One of ordinary skill in theart would be able to determine if a compound is capable of causingpositive inotropic effects and also additional beta-agonist compounds.In particular embodiments, the beta-receptor agonist is selective forthe beta-1 receptor. However, in other embodiments the beta-agonist isselective for the beta-2 receptor, or is not selective for anyparticular receptor.

Diseases or conditions that are “responsive to nitroxyl therapy” intendsany disease or condition in which administration of a compound thatdonates an effective amount of nitroxyl under physiological conditionstreats and/or prevents the disease or condition, as those terms aredefined herein. A disease or condition whose symptoms are suppressed ordiminished upon administration of nitroxyl donor is a disease orcondition responsive to nitroxyl therapy. Non-limiting examples ofdiseases or conditions that are responsive to nitroxyl therapy includecoronary obstructions, coronary artery disease (CAD), angina, heartattack, myocardial infarction, high blood pressure, ischemiccardiomyopathy and infarction, diastolic heart failure, pulmonarycongestion, pulmonary edema, cardiac fibrosis, valvular heart disease,pericardial disease, circulatory congestive states, peripheral edema,ascites, Chagas' disease, ventricular hypertrophy, heart valve disease,heart failure, including but not limited to congestive heart failuresuch as acute congestive heart failure and acute decompensated heartfailure. Other cardiovascular diseases or conditions are also intended,as are diseases or conditions that implicate ischemia/reperfusioninjury.

N-Hydroxysulfonamide Compounds

The compounds of this invention and for use in the methods describedherein include N-hydroxylsulfonamides that donate nitroxyl underphysiological conditions. Preferably, the compounds predominately donatenitroxyl under physiological conditions, meaning that a compound thatdonates both nitroxyl and nitric oxide under physiological conditionsdonates more nitroxyl than nitric oxide. Preferably, the compounds foruse herein do not donate significant levels of nitric oxide underphysiological conditions. Most preferably, the compounds for use hereindonate significant levels of nitroxyl under physiological conditions.

In one embodiment, the invention embraces a compound of the formula (I):

where R¹ is H; R² is H, aralkyl or heterocyclyl; R³, R⁴, R⁵, R⁶ and R⁷are independently selected from the group consisting of H, halo,alkylsulfonyl, N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl, cyano,alkoxy, perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl,alkylsulfinyl, heterocycloalkyl, substituted heterocycloalkyl,dialkylamino, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carboxyl, carboxyl ester, acylamino and sulfonylamino,provided that at least one of R³, R⁴, R⁵, R⁶ and R⁷ is carboxyl,carboxyl ester, acylamino or sulfonylamino.

In another embodiment, the compound is of the formula (I) where R¹ is H;R² is H, aralkyl or heterocyclyl; R⁴, R⁵ and R⁶ are independently H,halo, alkylsulfonyl, N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl,cyano, alkoxy, perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl,alkylsulfinyl, heterocycloalkyl, substituted heterocycloalkyl,dialkylamino, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carboxyl, carboxyl ester, acylamino or sulfonylamino; atleast one of R³ and R⁷ is an electron withdrawing group or a group thatsterically hinders the sulfonyl moiety, provided that at least one ofR³, R⁴, R⁵, R⁶ and R⁷ is carboxyl, carboxyl ester, acylamino orsulfonylamino. In one variation, at least one of R³ or R⁷ is an electronwithdrawing group. In another variation, both R³ and R⁷ are electronwithdrawing groups. In another variation, at least one of R³ or R⁷ is agroup that sterically hinders the sulfonyl moiety of compound (I). Inone variation, at least one of R³ or R⁷ is a branched alkyl group, suchas i-propyl or t-butyl. In another variation, both R³ and R⁷ are alkylgroups provided that one of the alkyl groups is a branched alkyl group,such as when both groups are isopropyl or when one group is ethyl andthe other is sec-butyl. In one variation, one of R³ and R⁷ is anelectron withdrawing group and the R³ or R⁷ that is not an electronwithdrawing group is an alkyl group, which may be a branched alkyl groupsuch as isopropyl.

Also embraced is a compound of the formula (I) where R¹ is H; R² is H,benzyl or tetrahydropyran-2-yl; R³, R⁴, R⁵, R⁶ and R⁷ are independentlyselected from the group consisting of H, Cl, F, I, Br, SO₂CH₃, SO₂NHOH,CF₃, NO₂, phenyl, CN, OCH₃, OCF₃, t-Bu, O-iPr,4-nitrophenyloxy(OPh4-NO₂), propane-2-thiyl(SCH(CH₃)₂),propane-2-sulfinyl (S(O)CH(CH₃)₂), morpholino, N-methyl-piperazino,dimethylamino, piperidino, cyclohexyloxy, cyclopentylsulfanyl,phenylsulfanyl, phenylsulfinyl, carboxyl, carboxyl ester, acylamino orsulfonylamino provided that at least one of R³, R⁴, R⁵, R⁶ and R⁷ iscarboxyl, carboxyl ester, acylamino or sulfonylamino.

For any of the variations described for formula (I), included arevariations of formula (I) where R¹ is H and R² is H, benzyl ortetrahydropyran-2-yl. In one variation, the compound is of the formula(I) where at least two of R³, R⁴, R⁵, R⁶ and R⁷ are halo, such as thecompound of formula (I) where R is halo (such as F or Br) and one of R³and R⁷ is halo (such as Br, or Cl) or where both R³ and R⁷ or both R³and R⁴ are halo (such as when both are Cl or both are F or one is Cl andone is F), and the remaining substituents are as described in thevariations above. In one variation, the compound is of the formula (I)where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —S(O)₂alkyl, such as whenone of R³ or R⁷ is —S(O)₂CH₃. In one variation, the compound is of theformula (I) where at least one of R³, R⁵ and R⁷ is a perhaloalkyl, suchas when R³ is CF₃ or when R³ and R⁵ are CF₃. In one variation, thecompound is of the formula (I) where R⁵ is CF₃ and at least one of R³and R⁷ is other than H, such as when R⁵ is CF₃ and R³ is NO₂ or Cl. Inone variation, the compound is of the formula (I) where at least one ofR³, R⁴, R⁵, R⁶ and R⁷ is an aryl group, such as when at least one of R³and R⁷ is an aryl group, such as phenyl. In one variation, the compoundis of the formula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is aheterocyclyl group, such as when at least one of R³, R⁵ and R⁷ is aheterocyclyl group or substituted heterocylco group, such as morpholino,N-methyl, piperizino and piperidino. In one variation, the compound isof the formula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is acycloalkoxy or cycloalkylsulfanyl group such as when at least one of R³,R⁵ and R⁷ is a cyclohexyloxy, cyclopentyloxy, cyclohexylsulfanyl orcyclopentylsulfanyl group. In one variation, the compound is of theformula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is anarylsulfanyl or arylsulfinyl group, such as when at least one of R³, R⁵and R⁷ is a phenylsulfanyl or phenylsulfinyl group.

For any of the variations described for formula (I), included arevariations of formula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ iscarboxyl. In one such variation, R⁴ is carboxyl, R³, R⁵ and R⁶ are H andR⁷ is H or halo. In a particular variation, R⁴ is carboxyl, R³, R⁵ andR⁶ are H, R⁷ is H or halo and R¹ and R² are H. For any of the variationsdescribed for formula (I), included are variations of formula (I) whereat least one of R³, R⁴, R⁵, R⁶ and R⁷ is —COO-alkyl. In one suchvariation, R³ is —COO-alkyl and R⁴, R⁵, R⁶ and R⁷ are H. In a particularvariation, R³ is —COO-alkyl, R⁴, R⁵, R⁶ and R⁷ are H and R¹ and R² areH. In another variation, R⁴ is —COO-alkyl, one of R⁶ and R⁷ is —SR¹¹,aryl, —OR¹¹, nitro, cyano, acyl, —S(O)₂NHOH, sulfonylamino, C₁-C₂perfluoroalkyl, lower alkyl or amino and the R⁶ or R⁷ that is not SR¹¹,aryl, —OR¹¹, nitro, cyano, acyl, —S(O)₂NHOH, sulfonylamino, C₁-C₂perfluoroalkyl, lower alkyl or amino is hydrogen. In another variation,R⁴ is —COO-alkyl, one of R⁶ and R⁷ is —SR¹¹, aryl, —OR¹¹, nitro, cyano,acyl, —S(O)₂NHOH, sulfonylamino, C₁-C₂ perfluoroalkyl, lower alkyl oramino and the R⁶ or R⁷ that is not —SR¹¹, aryl, —OR¹¹, nitro, cyano,acyl, —S(O)₂NHOH, sulfonamino, C₁-C₂ perfluoroalkyl, lower alkyl oramino is hydrogen, and R¹, R², R³ and R⁵ are hydrogen. For any of thevariations described for formula (I), included are variations of formula(I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —COO-substitutedalkyl. In one such variation, R⁴ is —COO-substituted alkyl, R³, R⁵ andR⁶ are H and R⁷ is halo. In a particular variation, R⁴ is—COO-substituted alkyl, R⁷ is halo, R³, R⁵ and R⁶ are H and R¹ and R²are H. For any of the variations described for formula (I), included arevariations of formula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is—C(O)NH₂. For any of the variations described for formula (I), includedare variations of formula (I) where at least one of R³, R⁴, R⁵, R⁶ andR⁷ is —C(O)NR_(a)R_(b) where R_(a) is hydrogen and R_(b) is alkyl. Inone such variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) is hydrogen,R_(b) is a lower alkyl, R³, R⁵ and R⁶ are H and R⁷ is halo. In aparticular variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) is hydrogen,R_(b) is lower alkyl, R³, R⁵ and R⁶ are H and R⁷ is halo and R¹ and R²are H. In a particular variation, R_(b) is a C₂-C₄ alkyl, such as ethyl,propyl or butyl. In another variation, R_(b) is a branched lower alkyl,e.g., isopropyl or isobutyl. For any of the variations described forformula (I), included are variations of formula (I) where at least oneof R³, R⁴, R⁵, R⁶ and R⁷ is —C(O)NR_(a)R_(b) where R_(a) is alkyl,substituted alkyl or hydrogen and R_(b) is substituted alkyl. In onesuch variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) is alkyl, substitutedalkyl or hydrogen, R_(b) is substituted alkyl, R³, R⁵ and R⁶ are H andR⁷ is halo. In a particular variation, R⁴ is —C(O)NR_(a)R_(b) whereR_(a) is alkyl, substituted alkyl or hydrogen, R_(b) is substitutedalkyl, R³, R⁵ and R⁶ are H and R⁷ is halo and R¹ and R² are H. In stillanother variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) is lower alkyl,substituted lower alkyl or hydrogen, R_(b) is substituted lower alkyl,R³, R⁵ and R⁶ are H and R⁷ is halo. When R_(b) is a substituted loweralkyl group, in one variation it is a lower alkyl substituted withhydroxyl, carboxyl, amino or alkoxy group. For example, the inventionembraces compounds where R⁴ is —C(O)NR_(a)R_(b) wherein R_(a) ishydrogen, methyl, ethyl, or a lower alkyl substituted with hydroxyl oralkoxy group, R_(b) is a lower alkyl substituted with hydroxyl,carboxyl, amino or alkoxy group, R³, R⁵ and R⁶ are H and R⁷ is halo; ina further variation R¹ and R² are H. For any of the variations describedfor formula (I), included are variations of formula (I) where at leastone of R³, R⁴, R⁵, R⁶ and R⁷ is —C(O)NR_(a)R_(b) where R_(a) and R_(b)are independently alkyl. In one such variation, R⁴ is —C(O)NR_(a)R_(b)where R_(a) and R_(b) are independently alkyl, R³, R⁵ and R⁶ are H andR⁷ is halo. In another variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) andR_(b) are independently alkyl, R³ and R⁵ are hydrogen and one of R⁶ andR⁷ is —SR¹¹, aryl, —OR¹¹, nitro, cyano, acyl, —S(O)₂NHOH, sulfonamino,C₁-C₂ perfluoroalkyl, lower alkyl or amino and the R⁶ or R⁷ that is not—SR¹¹, aryl, —OR¹¹, nitro, cyano, acyl, —S(O)₂NHOH, sulfonamino, C₁-C₂perfluoroalkyl, lower alkyl or amino is hydrogen. In a particularvariation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) and R_(b) areindependently alkyl, R³, R⁵ and R⁶ are H and R⁷ is halo and R¹ and R²are H. R_(a) and R_(b) may be the same or different, e.g., R_(a) andR_(b) in one variation are both methyl or ethyl. For any of thevariations described for formula (I), included are variations of formula(I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —C(O)NR_(a)R_(b)where R_(a) and R_(b) are taken together with the nitrogen to which theyare attached to form a heterocyclic or substituted heterocyclic ring. Inone such variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) and R_(b) aretaken together with the nitrogen to which they are attached to form aheterocyclic or substituted heterocyclic ring. In another variation, R⁴is —C(O)NR_(a)R_(b) where R_(a) and R_(b) are taken together with thenitrogen to which they are attached to form a heterocyclic orsubstituted heterocyclic ring, R³, R⁵ and R⁶ are H and R⁷ is halo. In aparticular variation, R⁴ is —C(O)NR_(a)R_(b) where R_(a) and R_(b) aretaken together with the nitrogen to which they are attached to form aheterocyclic or substituted heterocyclic ring, R³, R⁵ and R⁶ are H andR⁷ is halo and R¹ and R² are H. In one variation, R_(a) and R_(b) aretaken together with the nitrogen to which they are attached to form aheterocyclic ring, such as morpholino. In any variation herein where R⁴is —C(O)NR_(a)R_(b), in a particular variation, R_(a) and R_(b) aretaken together with the nitrogen to which they are attached to form aheterocyclic ring selected from piperazinyl, azetidinyl, pyrrolidinyl,piperidinyl, thiomorpholinyl and morpholinyl. In any variation hereinwhere R⁴ is —C(O)NR_(a)R_(b), in a particular variation, R_(a) and R_(b)are taken together with the nitrogen to which they are attached to forma heterocyclic ring substituted with 1 or 2 moieties selected from loweralkyl, carboxylester, acyl, halo, amino, hydroxyl, substituted loweralkyl, oxo and alkoxy. For example, in any variation herein where R⁴ is—C(O)NR_(a)R_(b), in a particular variation, R_(a) and R_(b) are takentogether with the nitrogen to which they are attached to form asubstituted heterocyclic ring selected from 2,6-dimethylpiperaz-4-yl,1-isopropylpiperaz-4-yl, 1-(piperazin-4-yl)ethanone, tert-butylpiperaz-4-yl-1-carboxylate, 4-fluoropiperidyl, 4,4-difluoropiperidyl,4-aminopiperidyl, 4-hydroxypiperidyl, 4-oxopiperidinyl,4-methoxypiperidyl, 4-(2-hydroxyethyl)piperidyl,2-(piperid-4-yl)-ethoxyethanol, 3-hydroxy-azetidinyl,2-oxo-piperazin-4-yl and 1-methyl-2-oxo-piperazin-4-yl. For any of thevariations described for formula (I), included are variations of formula(I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —SO₂NH₂. For any ofthe variations described for formula (I), included are variations offormula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —SO₂NR-alkylwhere R is hydrogen. For any of the variations described for formula(I), included are variations of formula (I) where at least one of R³,R⁴, R⁵, R⁶ and R⁷ is —SO₂NR— alkyl where R is alkyl. In a particularvariation, R⁴ is —SO₂NR-alkyl where R is alkyl and R³, R⁵, R⁶ and R⁷ arehydrogen. For example, in one variation, R⁴ is —SO₂N (lower alkyl)₂ andR³, R⁵, R⁶ and R⁷ are hydrogen, where the lower alkyl substituents maybe the same or different, e.g., R⁴ may be —SO₂N(Et)₂ or —SO₂N(Et)(Me).For any of the variations described for formula (I), included arevariations of formula (I) where at least one of R³, R⁴, R⁵, R⁶ and R⁷ is—SO₂NR₂, where the two R groups are taken together with the nitrogen towhich they are attached to form a heterocyclic or substitutedheterocyclic ring. In one such variation, R³ is —SO₂NR₂, where the two Rgroups are taken together with the nitrogen to which they are attachedto form a heterocyclic or substituted heterocyclic ring. In anothervariation, R³ is —SO₂NR₂, where the two R groups are taken together withthe nitrogen to which they are attached to form a heterocyclic orsubstituted heterocyclic ring and R⁴, R⁵, R⁶ and R⁷ are H. In aparticular variation, R³ is —SO₂NR₂, where the two R groups are takentogether with the nitrogen to which they are attached to form aheterocyclic or substituted heterocyclic ring and R⁴, R⁵, R⁶ and R⁷ areH and R¹ and R² are H. In one variation, at least one of R³, R⁴, R⁵, R⁶and R⁷ is —SO₂NR₂, where the two R groups are taken together with thenitrogen to which they are attached to form a heterocyclic ring, such asa morpholino ring.

Representative compounds of the formula (I) include, but are not limitedto, the compounds listed in Table 1.

TABLE 1 Representative Compounds of Formula (I):

Compound 1

Compound 2

Compound 3

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Compound 9

Compound 10

Compound 11

Compound 12

Compound 13

Compound 14

Compound 15

Compound 16

Compound 17

Compound 18

Compound 19

Compound 20

Compound 21

Compound 22

Compound 23

Compound 24

Compound 25

Compound 26

Compound 27

Compound 28

Compound 29

Compound 30

Compound 31

Compound 32

Compound 33

Compound 34

Compound 35

Compound 36

Compound 37

Compound 38

Compound 39

Compound 40

Compound 41

Compound 42

Compound 43

Compound 44

Compound 45

Compound 46

Compound 47

Compound 48

Compound 49

Compound 50

Compound 51

Compound 52

Compound 53

Compound 54

Compound 55

Compound 56

Compound 57

Compound 58

Compound 59

Compound 60

Compound 61

Compound 62

Compound 63

Compound 64

Compound 65

Compound 66

Compound 67

Compound 68

Compound 69

Compound 70

Compound 71

Compound 72

In one embodiment, the nitroxyl donating compound is a compound of theformula (IIa):

where R¹ is H; R² is H, aralkyl or heterocyclyl; m and n areindependently an integer from 0 to 1; x is an integer from 0 to 4 and yis an integer from 0 to 3, provided that at least one of x and y isgreater than 0; A is a cycloalkyl, heterocycloalkyl, aromatic orheteroaromatic ring containing ring moieties Q¹, Q², Q³ and Q⁴, whichare taken together with the carbons at positions a and a′ to form ringA; B is a cycloalkyl, heterocycloalkyl, aromatic or heteroaromatic ringcontaining ring moieties Q⁵, Q⁶, Q⁷ and Q⁸, which are taken togetherwith the carbons at positions a and a′ to form ring B; Q¹, Q², Q³, Q⁴,Q⁵, Q⁶, Q⁷ and Q⁸ are independently selected from the group consistingof C, CH₂, CH, N, NR¹⁰, O and S; each R⁸ and R⁹ is independentlyselected from the group consisting of halo, alkylsulfonyl,N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl, cyano, alkoxy,perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl, alkylsulfinyl,heterocycloalkyl, substituted heterocycloalkyl, dialkylamino, NH₂, OH,C(O)OH, C(O)Oalkyl, NHC(O)alkylC(O)OH, C(O)NH₂, NHC(O)alkylC(O)alkyl,NHC(O)alkenylC(O)OH, NHC(O)NH₂, OalkylC(O)Oalkyl, NHC(O)alkyl,C(═N—OH)NH₂, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carbonylamino and sulfonylamino, provided that at leastone R⁸ is carbonylamino or sulfonylamino; and, R¹⁰ is H, alkyl, acyl, orsulfonyl.

In one variation, the compound is of the formula (II) or (Ia) where eachR⁸ and R⁹ is independently selected from the group consisting of Cl, F,I, Br, SO₂CH₃, SO₂NHOH, CF₃, CH₃, NO₂, phenyl, CN, OCH₃, OCF₃, t-Bu,O-iPr, 4-nitrophenyloxy(OPh4-NO₂), propane-2-thiyl(SCH(CH₃)₂),propane-2-sulfinyl(S(O)CH(CH₃)₂), morpholino, N-methyl-piperazino,dimethylamino, piperidino, cyclohexyloxy, cyclopentylsulfanyl,phenylsulfanyl, phenylsulfinyl, carbonylamino and sulfonylamino,provided that at least one R⁸ is carbonylamino or sulfonylamino; and R¹⁰is H, alkyl, acyl or sulfonyl, provided that when rings A and B formnaphthalene, x is an integer from 1 to 3 or y is an integer from 2 to 4.

For any of the variations described for formula (II) or (Ia), includedare variations of formula (II) or (IIa) where R¹ is H and R² is H,benzyl or tetrahydropyran-2-yl. In one variation, A and B form abenzofuran or benzothiophene or benzoimidazole or N-alkylbenzoimidazole(such as N-methylbenzoimidazole) or N-acylbenzoimidazole (such asN—C(O)CH₃benzoimidazole) or benzothiazole or benzooxazole. In onevariation, A and B are other than napthyl or quinoline. In onevariation, A and B are napthyl or quinoline. In one variation, A and Bform a benzofuran. In one variation, A and B form a benzofuran. In onevariation, A and B form a benzothiophene. In one variation, A and B forma benzothiophene, y is 0 and x is 1. In one variation, A and B formnaphthyl and x is 0, y is 1. In one variation, ring A is phenyl and ringB is a heteroaryl group, such as when rings A and B form quinoline andring B is the nitrogen containing ring. The invention also embracescompounds according to any of the variations for formula (II) or (IIa)where y is 0, x is 1 and R⁹ is a halo, alkyl or perhaloalkyl group. Theinvention also embraces compounds according to any of the variations forformula (II) or (IIa) where x is 2 and y is 0.

For any of the variations described for formula (II) or (Ia), includedare variations of formula (II) or (IIa) where at least one of R⁸ and R⁹is —CONH-alkyl. For any of the variations described for formula (II) or(IIa), included are variations of formula (II) or (IIa) where at leastone of R⁸ and R⁹ is —CONR-alkyl where R is alkyl. For any of thevariations described for formula (II) or (IIa), included are variationsof formula (II) or (Ia) where at least one of R⁸ and R⁹ is —CONR₂ whereeach R is independently alkyl. In a particular variation of formula (II)or (IIa), y is 0, x is 1 and R⁹ is —CONR₂ where each R is independentlyalkyl. In another variation of formula (II) or (IIa), y is 0, x is 1 andR⁹ is —CONR₂ where each R is independently lower alkyl, where each loweralkyl can be the same (e.g., —CON(Me)₂) or different. For any of thevariations described for formula (II) or (Ia), included are variationsof formula (II) or (IIa) where at least one of R⁸ and R⁹ is —CONR₂ whereeach R taken together with the nitrogen to which it is attached to forma heterocylic or substituted heterocyclic ring. For any of thevariations described for formula (II) or (IIa), included are variationsof formula (II) or (IIa) where at least one of R⁸ and R⁹ is —CONR₂ whereeach R is independently alkyl. For any of the variations described forformula (II) or (IIa), included are variations of formula (II) or (IIa)where at least one of R⁸ and R⁹ is —NR^(a)SO₂NR-alkyl where R_(a) and Rare independently hydrogen or alkyl. For any of the variations describedfor formula (II) or (Ia), included are variations of formula (II) or(IIa) where at least one of R⁸ and R⁹ is —SO₂NH₂. For any of thevariations described for formula (II) or (IIa), included are variationsof formula (II) or (IIa) where at least one of R⁸ and R⁹ is —SO₂NH₂. Forany of the variations described for formula (II) or (IIa), included arevariations of formula (II) or (IIa) where at least one of R⁸ and R⁹ is—SO₂NR-alkyl, where R is hydrogen or alkyl. For any of the variationsdescribed for formula (II) or (IIa), included are variations of formula(II) or (IIa) where at least one of R⁸ and R⁹ is —SO₂NR₂, where the twoR groups are taken together and with the nitrogen atom to which they areattached to form a heterocyclic or substituted heterocyclic ring. In aparticular variation of formula (II) or (IIa), y is 0, x is 1 and R⁹ is—SO₂NR₂, where the two R groups are taken together and with the nitrogenatom to which they are attached to form a heterocyclic ring. In anothervariation of formula (II) or (IIa), y is 0, x is 1 and R⁹ is —SO₂NR₂,where the two R groups are taken together and with the nitrogen atom towhich they are attached to form a morpholino group.

Representative compounds of the formula (IIa) include, but are notlimited to, the compounds listed in Table 2.

TABLE 2 Representative Compounds of Formula (IIa):

Compound 73

Compound 74

Compound 75

Compound 76

In another embodiment, the nitroxyl donating compound is a compound ofthe formula (III):

where R¹ is H; R² is H, aralkyl or heterocyclyl; n is an integer from 0to 1; b is an integer from 1 to 4; C is a heteroaromatic ring containingring moieties Q⁹, Q¹⁰, Q¹¹, Q¹², Q¹³ and Q¹⁴ that are independentlyselected from the group consisting of C, CH₂, CH, N, NR¹⁰, O and S,provided that at least one of Q⁹, Q¹⁰, Q¹¹, Q¹², Q¹³ and Q¹⁴ is N, NR¹⁰,O or S; each R⁸ is independently selected from the group consisting ofhalo, alkylsulfonyl, N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl,cyano, alkoxy, perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl,alkylsulfinyl, heterocycloalkyl, substituted heterocycloalkyl,dialkylamino, NH₂, OH, C(O)OH, C(O)Oalkyl, NHC(O)alkylC(O)OH, C(O)NH₂,NHC(O)alkylC(O)alkyl, NHC(O)alkenylC(O)OH, NHC(O)NH₂, OalkylC(O)Oalkyl,NHC(O)alkyl, C(═N—OH)NH₂, cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl,arylsulfinyl, carbonylamino or sulfonylamino, provided that at least oneR⁸ is carbonylamino or sulfonylamino; and R¹⁰ is H, alkyl, acyl orsulfonyl.

In one variation, the compound is of the formula (III) and each R⁸ isindependently selected from the group consisting of Cl, F, I, Br,SO₂CH₃, SO₂NHOH, CF₃, CH₃, NO₂, phenyl, CN, OCH₃, OCF₃, t-Bu, O-iPr,4-nitrophenyloxy(OPh4-NO₂), propane-2-thiyl (SCH(CH₃)₂),propane-2-sulfinyl(S(O)CH(CH₃)₂), morpholino, N-methyl-piperazino,dimethylamino, piperidino, cyclohexyloxy, cyclopentylsulfanyl,phenylsulfanyl, phenylsulfinyl, carbonylamino or sulfonylamino, providedthat at least one R⁸ is carbonylamino or sulfonylamino. In anothervariation, the compound is of the formula (III) and each R⁸ isindependently selected from the group consisting of F, Br, C₁, CF₃,phenyl, methyl, SO₂NHOH, morpholino, piperidino, 4-methyl-piperazino,carbonylamino and sulfonylamino, provided that at least one R⁸ iscarbonylamino or sulfonylamino.

For any of the variations described for formula (III), included arevariations of formula (III) where R¹ is H and R² is H, benzyl ortetrahydropyran-2-yl. In one variation, n is 0 and C is a thiophene orisoxazole or pyrazole or pyrrole or imidazole or furan or thiazole ortriazole or N-methylimidazole or thiadiazole. In one variation, C isother than thienyl. In another variation, n is 0 and C is a thiophene orisoxazole or pyrazole or pyrrole or imidazole or furan or thiazole ortriazole or N-methylimidazole or thiadiazole. In one variation, n is 1and C is a pyrimidine or pyrazine or pyridine. In one variation, n is 1and C is a pyrimidine or pyrazine or pyridine and b is 1. In onevariation, n is 1 and C is a pyrimidine or pyrazine or pyridine, b is 1,and at least one R⁸ is chloro or morpholino or piperidino orN-methylpiperizino. In one variation, C is thiophene and b is 1. In onevariation, C is thiophene, b is 1 and at least one R⁸ is halo. In onevariation, C is thiophene.

For any of the variations described for formula (III), included arevariations of formula (III) where at least one R⁸ is —CONH₂. In onevariation, C is a thiophene substituted with —CONH₂, and optionallysubstituted with an additional R⁸, such as an amino group. For any ofthe variations described for formula (III), included are variations offormula (III) where at least one R⁸ is —CONH-alkyl. For example,compounds wherein R⁸ is —CONH-lower alkyl (e.g., isopropyl) areencompassed. For any of the variations described for formula (III),included are variations of formula (III) where at least one R⁸ is—CONH-substituted alkyl. In a particular variation, C is thiophene, R¹and R² are both H and at least one R¹ is —CONH-substituted alkyl. Forany of the variations described for formula (III), included arevariations of formula (III) where at least one R⁸ is —CONR-alkyl where Ris alkyl. For any of the variations described for formula (III),included are variations of formula (III) where at least one R⁸ is —CONR₂where each R is independently alkyl, such as —CON(Me)₂. In a particularvariation, C is a thiophene, b is 2, one of R⁸ is —CONR₂ where each R isindependently alkyl (such as —CON(Me)₂) and the other R⁸ is —S(O)₂Alkyl,aryl, heteroaryl, or —S-alkyl. For any of the variations described forformula (III), included are variations of formula (III) where at leastone R⁸ is —CONR₂ where each R is independently a substituted alkyl, suchas —CH₂CH₂OCH₃. For any of the variations described for formula (III),included are variations of formula (III) where at least one R¹ is —CONR₂where each R taken together with the nitrogen to which it is attached toform a heterocylic or substituted heterocyclic ring. In a particularvariation, C is thiophene, R¹ and R² are both H and at least one R⁸ is—CONR₂ where each R taken together with the nitrogen to which it isattached to form a heterocylic or substituted heterocyclic ring, such asmorpholino. In a another variation, C is thiophene, R¹ and R² are bothH, b is 1 or 2, at least one R¹ is —CONR₂ where each R taken togetherwith the nitrogen to which it is attached to form a heterocylic ringselected from piperidinyl and morpholinyl and when b is 2, the R⁸ thatis other than is —CONR₂ is selected from halo, nitro and —OR¹¹, such as—Oalkyl (e.g., methoxy). In a another variation, C is thiophene, R¹ andR² are both H, b is 1 or 2, at least one R⁸ is —CONR₂ where each R takentogether with the nitrogen to which it is attached to form aheterocyclic ring substituted with 1 or 2 moieties selected from loweralkyl, carboxylester, acyl, halo, amino, hydroxyl, substituted loweralkyl, oxo and alkoxy. For example, in any variation herein where R⁸ is—CONR₂ where each R taken together with the nitrogen to which it isattached to form a substituted heterocyclic ring selected from1-methyl-piperaz-4-yl, 4-fluoropiperidyl and 4-hydroxypiperidyl. For anyvariation, e.g., when C is thiophene substituted with R⁸ is —CONR₂, Cmay also be substituted with a moiety selected from halo, amino,hydroxyl, alkoxy, nitro and cyano. For any of the variations describedfor formula (III), included are variations of formula (III) where atleast one R⁸ is —CONR₂ where each R is independently alkyl. For any ofthe variations described for formula (III), included are variations offormula (III) where at least one R⁸ is —NR^(a)SO₂NR-alkyl where R_(a)and R are independently hydrogen or alkyl. For any of the variationsdescribed for formula (III), included are variations of formula (III)where at least one R⁸ is —SO₂NH₂. For any of the variations describedfor formula (III), included are variations of formula (III) where atleast one R⁸ is —SO₂NH₂. For any of the variations described for formula(III), included are variations of formula (III) where at least one R⁸ is—SO₂NR-alkyl, where R is hydrogen or alkyl. For any of the variationsdescribed for formula (III), included are variations of formula (III)where at least one R⁸ is —SO₂NR₂, where the two R groups are takentogether and with the nitrogen atom to which they are attached to form aheterocyclic or substituted heterocyclic ring.

Representative compounds of the formula (III) include, but are notlimited to, the compounds listed in Table 3.

TABLE 3 Representative Compounds of Formula (III):

Compound 97

Compound 98

Compound 77

Compound 78

Compound 79

Compound 80

Compound 81

Compound 82

Compound 83

Compound 84

Compound 85

Compound 86

Compound 87

Compound 88

Compound 89

Compound 90

Compound 91

Compound 92

Compound 93

Compound 94

Compound 95

Compound 96Compounds for Use in the Methods

The methods described employ compounds of the invention that donate aneffective amount of nitroxyl under physiological conditions. Any of themethods may employ an N-hydroxylsulfonamide compound described aboveunder “N-Hydroxysulfonamide Compounds.”

For any of the compounds of the invention, such as the compounds offormula (I), (II) or (III) or other compounds for use in the methodsdescribed herein, recitation or depiction of the parent compound intendsand includes all salts, solvates, hydrates, polymorphs, or prodrugsthereof, where applicable. As such, all salts, such as pharmaceuticallyacceptable salts, solvates, hydrates, polymorphs and prodrugs of acompound are embraced by the invention and described herein the same asif each and every salts, solvate, hydrate, polymorph, or prodrug werespecifically and individually listed.

For all compounds disclosed herein, where applicable due to the presenceof a stereocenter, the compound is intended to embrace all possiblestereoisomers of the compound depicted or described. Compositionscomprising a compound with at least one stereocenter are also embracedby the invention, and includes racemic mixtures or mixtures containingan enantiomeric excess of one enantiomer or single diastereomers ordiastereomeric mixtures. All such isomeric forms of these compounds areexpressly included herein the same as if each and every isomeric formwere specifically and individually listed. The compounds herein may alsocontain linkages (e.g., carbon-carbon bonds) wherein bond rotation isrestricted about that particular linkage, e.g. restriction resultingfrom the presence of a ring or double bond. Accordingly, all cis/transand E/Z isomers are also expressly included in the present invention.The compounds herein may also be represented in multiple tautomericforms, in such instances, the invention expressly includes alltautomeric forms of the compounds described herein, even though only asingle tautomeric form may be represented. Also embraced arecompositions of substantially pure compound. A composition ofsubstantially pure compound means that the composition contains no morethan 25%, or no more than 15%, or no more than 10%, or no more than 5%,or no more than 3% impurity, or no more than 1% impurity, such as adifferent biologically active compound, which may include a differentstereochemical form of the compound if the composition contains asubstantially pure single isomer.

The compounds of the invention can be made according to the generalmethods described in Schemes A-D or by procedures known in the art.Starting materials for the reactions are either commercially availableor may be prepare by known procedures or obvious modifications thereof.For example, many of the starting materials are available fromcommercial suppliers such as Sigma-Aldrich. Others may be prepared byprocedures or obvious modifications thereof described in standardreference texts such as March's Advanced Organic Chemistry, (John Wileyand Sons) and Larock's Comprehensive Organic Transformations (VCHPublishers Inc.).

In Scheme A, a solution of hydroxylamine hydrochloride in water ischilled to 0° C. A solution of potassium carbonate in water is addeddropwise, maintaining an internal reaction temperature between about 5°C. and about 15° C. The reaction mixture is stirred for about 15minutes, whereupon tetrahydrofuran (THF) and methanol (MeOH) are added.Compound A1 (where R is an alkyl, aryl or heterocyclyl group) is addedportionwise maintaining a temperature below about 15° C. and thereaction mixture is stirred at ambient temperature until completeconsumption of the sulfonyl chloride is observed by thin layerchromatography (TLC). The resulting suspension is concentrated to removeany volatiles and the aqueous suspension is extracted with diethylether. The organic portion is dried over magnesium sulfate, filtered andconcentrated in vacuo to yield the crude N-hydroxy sulphonamide A2.Purification may be achieved by conventional methods, such aschromatography, filtration, crystallization and the like.

In Scheme B, a solution of aqueous hydroxylamine in water and THF ischilled to −5° C. Compound A1 (where R is an alkyl, aryl or heterocyclylgroup) is added portionwise maintaining a temperature below about 10° C.and the reaction mixture is stirred at ambient temperature untilcomplete consumption of the sulfonyl chloride is observed by thin layerchromatography (TLC). The resulting suspension is concentrated to removeany volatiles and the aqueous suspension is extracted with diethylether. The organic portion is dried over magnesium sulfate, filtered andconcentrated in vacuo to yield the crude N-hydroxy sulphonamide A2.Purification may be achieved by conventional methods, such aschromatography, filtration, crystallization and the like.

N-Benzyloxysulfonamides are chemical intermediates that are used asprotected N-hydroxysulfonamides for the further modification of the Rmoiety of compound B2. In Scheme B, a suspension ofO-benzylhydroxylamine hydrochloride B1 in methanol and water is added toa chilled solution of potassium carbonate in water, maintaining aninternal reaction temperature below about 10° C. The reaction mixture isstirred for about 5 minutes, whereupon THF and A1 (where R is an alkyl,aryl or heterocyclyl group) are added. The reaction mixture is stirredat ambient temperature until complete consumption of the sulfonylchloride was observed by TLC. The resulting suspension is concentratedin vacuo to remove any volatiles, and the aqueous suspension wasextracted with diethyl ether. The organic layer was dried over sodiumsulfate, filtered and concentrated in vacuo to yield the crude targetcompound B2. Purification may be achieved by conventional methods, suchas chromatography, filtration, crystallization and the like. Thereaction product B2 may be deprotected by removing the benzyl group. Forinstance, a suspension of 10% palladium on charcoal may be added to asuspension of B2 in methanol. The reaction mixture is stirred under ahydrogen atmosphere at ambient temperature and atmospheric pressureovernight. The reaction mixture is filtered through microfibre glasspaper. The resulting filtrate is concentrated in vacuo, and the residuepurified by conventional methods to yield the correspondingN-hydroxylsulfonamide.

N-(tetrahydro-pyran-2-yloxy)sulfonamides are chemical intermediates thatare used as protected N-hydroxysulfonamides for the further modificationof the R moiety of compound C2. In Scheme C, to a solution of C1 inwater at 0° C. is added a solution of potassium carbonate in waterdropwise, maintaining an internal reaction temperature below about 10°C. After about 15 minutes, methanol and THF are added dropwise, followedby A1 portionwise. The reaction mixture is stirred at ambienttemperature until complete consumption of the sulfonyl chloride isobserved by TLC. The resulting suspension was concentrated to remove anyvolatiles and the aqueous suspension was extracted with diethyl ether.The organic portion is dried over sodium sulfate, filtered andconcentrated in vacuo to yield the crude target compound C2.Purification may be achieved by conventional methods, such aschromatography, filtration, crystallization and the like. Deprotectionof C2 to yield the corresponding N-hydroxylsulfonamide may be carriedout according to methods known in the art.

Methods of Using the Compounds and Compositions

The compounds and compositions herein may be used to treat and/orprevent the onset and/or development of a disease or condition that isresponsive to nitroxyl therapy.

The invention embraces methods of administering to an individual(including an individual identified as in need of such treatment) aneffective amount of a compound to produce a desired effect. Identifyinga subject in need of such treatment can be in the judgment of aphysician, clinical staff, emergency response personnel or other healthcare professional and can be subjective (e.g. opinion) or objective(e.g. measurable by a test or diagnostic method).

One embodiment provides a method of modulating (including increasing) invivo nitroxyl levels in an individual in need thereof, the methodcomprising administering to the individual a compound that donatesnitroxyl under physiological conditions or a pharmaceutically acceptablesalt thereof. An individual is in need of nitroxyl modulation if theyhave or are suspected of having or are at risk of having or developing adisease or condition that is responsive to nitroxyl therapy.

Particular diseases or conditions embraced by the methods of theinvention include cardiovascular diseases such as heart failure orconditions and diseases or conditions that implicate or may implicateischemia/reperfusion injury. These methods are described in more detailbelow.

Compositions comprising a nitroxyl-donating compound of the inventionare embraced by the invention. However, the methods described may usemore than one nitroxyl donating compound; for example, the methods mayemploy Angeli's salt and an N-hydroxysulfonamide of the presentinvention or two or more N-hydroxysulfonamides of the present invention,which may be administered together or sequentially.

Cardiovascular Diseases

Provided herein are methods of treating cardiovascular diseases such asheart failure by administering an effective amount of at least onenitroxyl donating compound to an individual in need thereof. Alsoprovided are methods of administering a therapeutically effective doseof at least one nitroxyl donating compound in combination with at leastone other positive inotropic agent to an individual in need thereof.Further provided are methods of administering a therapeuticallyeffective amount of at least one nitroxyl donating compound to anindividual who is receiving beta-antagonist therapy and who isexperiencing heart failure. Methods are provided herein foradministering compounds of the invention in combination withbeta-adrenergic agonists to treat heart failure. Such agonists includedopamine, dobutamine, and isoproterenol, and analogs and derivatives ofsuch compounds. Also provided are methods of administering nitroxyldonors to individuals receiving treatment with beta-antagonizing agentssuch as propranolol, metoprolol, bisoprolol, bucindolol, and carvedilol.Further, methods are provided herein for treating specificclassifications of heart failure, such as Class III heart failure andacute heart failure.

Also embraced by the invention is a method of treating congestive heartfailure (CHF), including acute congestive heart failure, byadministering an effective amount at least one nitroxyl donatingcompound to an individual in need thereof, which individual may beexperiencing heart failure. Also disclosed is a method of treating CHFby administering an effective amount of at least one nitroxyl donatingcompound in combination with an effective amount of at least one otherpositive inotropic agent to an individual in need thereof, whichindividual may be experiencing heart failure. In one variation, theother positive inotrope is a beta-adrenergic agonist, such asdobutamine. The combined administration of a nitroxyl donor and at leastone other positive inotropic agent comprises administering the nitroxyldonor either sequentially with the other positive inotropic agent forexample, the treatment with one agent first and then the second agent,or administering both agents at substantially the same time, whereinthere is an overlap in performing the administration. With sequentialadministration, an individual is exposed to the agents at differenttimes, so long as some amount of the first agent, which is sufficient tobe therapeutically effective in combination with the second agent,remains in the subject when the other agent is administered. Treatmentwith both agents at the same time can involve administration of theagents in the same dose, such as a physically mixed dose, or in separatedoses administered at the same time.

In particular an embodiment, a nitroxyl donor is administered to anindividual experiencing heart failure that is receiving beta-antagonisttherapy. A beta-antagonist (also known as a beta-blocker) includes anycompound that effectively acts as an antagonist at a subject'sbeta-adrenergic receptors, and provides desired therapeutic orpharmaceutical results, such as diminished vascular tone and/or heartrate. A subject who is receiving beta-antagonist therapy is any subjectto whom a beta-antagonist has been administered, and in whom thebeta-antagonist continues to act as an antagonist at the subject'sbeta-adrenergic receptors. In particular embodiments a determination ofwhether a subject is receiving beta-blocking therapy is made byexamination of the subject's medical history. In other embodiments thesubject is screened for the presence of beta-blocking agents by chemicaltests, such as high-speed liquid chromatography as described in Theviset al., Biomed. Chromatogr., 15:393-402 (2001).

The administration of a nitroxyl donating compound either alone, incombination with a positive inotropic agent, or to a subject receivingbeta-antagonist therapy, is used to treat heart failure of allclassifications. In particular embodiments a nitroxyl donating compoundis used to treat early-stage chronic heart failure, such as Class IIheart failure. In other embodiments a nitroxyl donating compound is usedin combination with a positive inotropic agent, such as isoproterenol totreat Class IV heart failure. In still other embodiments a nitroxyldonating compound is used in combination with another positive inotropicagent, such as isoproterenol to treat acute heart failure. In someembodiments, when a nitroxyl donor is used to treat early stage heartfailure, the dose administered is lower than that used to treat acuteheart failure. In other embodiments the dose is the same as is used totreat acute heart failure.

Ischemia/Reperfusion Injury

The invention embraces methods of treating or preventing or protectingagainst ischemia/reperfusion injury. In particular, compounds of theinvention are beneficial for individuals at risk for an ischemic event.Thus, provided herein is a method of preventing or reducing the injuryassociated with ischemia/reperfusion by administering an effectiveamount of at least one nitroxyl donating compound to an individual,preferably prior to the onset of ischemia. A compound of the inventionmay be administered to an individual after ischemia but beforereperfusion. A compound of the invention may also be administered afterischemia/reperfusion, but where the administration protects againstfurther injury. Also provided is a method in which the individual isdemonstrated to be at risk for an ischemic event. Also disclosed is amethod of administering a nitroxyl donating compound to an organ that isto be transplanted in an amount effective to reduce ischemia/reperfusioninjury to the tissues of the organ upon reperfusion in the recipient ofthe transplanted organ.

Nitroxyl donors of the invention may thus be used in methods ofpreventing or reducing injury associated with futureischemia/reperfusion. For example, administration of a nitroxyl donorprior to the onset of ischemia may reduce tissue necrosis (the size ofinfarct) in at-risk tissues. In live subjects this may be accomplishedby administering an effective amount of a nitroxyl donating compound toan individual prior to the onset of ischemia. In organs to betransplanted this is accomplished by contacting the organ with anitroxyl donor prior to reperfusion of the organ in the transplantrecipient. Compositions comprising more than one nitroxyl-donatingcompound also could be used in the methods described, for example,Angeli's salt and an N-hydroxysulfonamide of the present invention ortwo or more N-hydroxysulfonamides of the present invention. Thenitroxyl-donating compound also can be used in combination with otherclasses of therapeutic agents that are designed to minimize ischemicinjury, such as beta blockers, calcium channel blockers, anti-platelettherapy or other interventions for protecting the myocardium inindividuals with coronary artery disease.

One method of administering a nitroxyl donor to live subjects includesadministration of the nitroxyl-donating compound prior to the onset ofischemia. This refers only to the onset of each instance of ischemia andwould not preclude performance of the method with subjects who have hadprior ischemic events, i.e., the method also contemplates administrationof nitroxyl-donating compounds to a subject who has had an ischemicevent in the past.

Individuals can be selected who are at risk of a first or subsequentischemic event. Examples include individuals with knownhypercholesterolemia, EKG changes associated with risk of ischemia,sedentary lifestyle, angiographic evidence of partial coronary arteryobstruction, echocardiographic evidence of myocardial damage, or anyother evidence of a risk for a future or additional ischemic event (forexample a myocardial ischemic event, such as a myocardial infarction(MI), or a neurovascular ischemia such as a cerebrovascular accidentCVA). In particular examples of the methods, individuals are selectedfor treatment who are at risk of future ischemia, but who have nopresent evidence of ischemia (such as electrocardiographic changesassociated with ischemia (for example, peaked or inverted T-waves or STsegment elevations or depression in an appropriate clinical context),elevated CKMB, or clinical evidence of ischemia such as crushingsub-sternal chest pain or arm pain, shortness of breath and/ordiaphoresis). The nitroxyl-donating compound also could be administeredprior to procedures in which myocardial ischemia may occur, for examplean angioplasty or surgery (such as a coronary artery bypass graftsurgery). Also embraced is a method of administering a nitroxyl-donatingcompound to an individual at demonstrated risk for an ischemic event.The selection of an individual with such a status could be performed bya variety of methods, some of which are noted above. For example, anindividual with one of more of an abnormal EKG not associated withactive ischemia, prior history of myocardial infarction, elevated serumcholesterol, etc., would be at risk for an ischemic event. Thus, anat-risk individual could be selected by physical testing or elicitingthe potential subject's medical history to determine whether the subjecthas any indications of risk for an ischemic event. If risk isdemonstrated based on the indications discussed above, or any otherindications that one skilled in the art would appreciate, then theindividual would be considered at demonstrated risk for an ischemicevent.

Ischemia/reperfusion may damage tissues other than those of themyocardium and the invention embraces methods of treating or preventingsuch damage. In one variation, the method finds use in reducing injuryfrom ischemia/reperfusion in the tissue of the brain, liver, gut,kidney, bowel, or in any other tissue. The methods preferably involveadministration of a nitroxyl donor to an individual at risk for suchinjury. Selecting a person at risk for non-myocardial ischemia couldinclude a determination of the indicators used to assess risk formyocardial ischemia. However, other factors may indicate a risk forischemia/reperfusion in other tissues. For example, surgery patientsoften experience surgery related ischemia. Thus, individuals scheduledfor surgery could be considered at risk for an ischemic event. Thefollowing risk factors for stroke (or a subset of these risk factors)would demonstrate a subject's risk for ischemia of brain tissue:hypertension, cigarette smoking, carotid artery stenosis, physicalinactivity, diabetes mellitus, hyperlipidemia, transient ischemicattack, atrial fibrillation, coronary artery disease, congestive heartfailure, past myocardial infarction, left ventricular dysfunction withmural thrombus, and mitral stenosis. Ingall, “Preventing ischemicstroke: current approaches to primary and secondary prevention,”Postgrad. Med., 107(6):34-50 (2000). Further, complications of untreatedinfectious diarrhea in the elderly can include myocardial, renal,cerebrovascular and intestinal ischemia. Slotwiner-Nie & Brandt,“Infectious diarrhea in the elderly,” Gastroenterol, Clin. N. Am.,30(3):625-635 (2001). Alternatively, individuals could be selected basedon risk factors for ischemic bowel, kidney or liver disease. Forexample, treatment would be initiated in elderly subjects at risk ofhypotensive episodes (such as surgical blood loss). Thus, subjectspresenting with such an indication would be considered at risk for anischemic event. Also embraced is a method of administering a nitroxyldonating compound of the invention to an individual who has any one ormore of the conditions listed herein, such as diabetes mellitus orhypertension. Other conditions that may result in ischemia such ascerebral arteriovenous malformation would be considered to demonstraterisk for an ischemic event.

The method of administering nitroxyl to organs to be transplantedincludes administration of nitroxyl prior to removal of the organ fromthe donor, for example through the perfusion cannulas used in the organremoval process. If the organ donor is a live donor, for example akidney donor, the nitroxyl donor can be administered to the organ donoras described above for a subject at risk for an ischemic event. In othercases the nitroxyl donor can be administered by storing the organ in asolution comprising the nitroxyl donor. For example, the nitroxyl donorcan be included in the organ preservation solution, such as Universityof Wisconsin “UW” solution, which is a solution comprising hydroxyethylstarch substantially free of ethylene glycol, ethylene chlorohydrin andacetone (see U.S. Pat. No. 4,798,824).

Pharmaceutical Composition, Dosage Forms and Treatment Regimens

Also included are pharmaceutically acceptable compositions comprising acompound of the invention or pharmaceutically acceptable salt thereofand any of the methods may employ the compounds of the invention as apharmaceutically acceptable composition. A pharmaceutically acceptablecomposition includes one or more of the compounds of the inventiontogether with a pharmaceutically acceptable carrier. The pharmaceuticalcompositions of the invention include those suitable for oral, rectal,nasal, topical (including buccal and sublingual), vaginal or parenteral(including subcutaneous, intramuscular, intravenous and intradermal)administration.

The compounds or compositions may be prepared as any available dosageform. Unit dosage forms are also intended, which includes discrete unitsof the compound or composition such as capsules, sachets or tablets eachcontaining a predetermined amount of the compound; as a powder orgranules; as a solution or a suspension in an aqueous liquid or anon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion, or packed in liposomes and as a bolus,etc.

A tablet containing the compound or composition may be made bycompression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with a binder, lubricant, inertdiluent, preservative, surface-active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsoptionally may be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient therein.Methods of formulating such slow or controlled release compositions ofpharmaceutically active ingredients, such as those herein and othercompounds known in the art, are known in the art and described inseveral issued U.S. patents, some of which include, but are not limitedto, U.S. Pat. Nos. 4,369,174 and 4,842,866, and references citedtherein. Coatings can be used for delivery of compounds to the intestine(see, e.g., U.S. Pat. Nos. 6,638,534, 5,217,720 and 6,569,457, andreferences cited therein). A skilled artisan will recognize that inaddition to tablets, other dosage forms can be formulated to provideslow or controlled release of the active ingredient. Such dosage formsinclude, but are not limited to, capsules, granulations and gel-caps.

Compositions suitable for topical administration include lozengescomprising the ingredients in a flavored basis, usually sucrose andacacia or tragacanth; and pastilles comprising the active ingredient inan inert basis such as gelatin and glycerin, or sucrose and acacia.

Compositions suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. The formulations may be presented in unit-dose or multi-dosecontainers, for example, sealed ampules and vials, and may be stored ina freeze dried (lyophilized) condition requiring only the addition ofthe sterile liquid carrier, for example water for injections,immediately prior to use.

Extemporaneous injection solutions and suspensions may be prepared fromsterile powders, granules and tablets.

Administration of the compounds or compositions to an individual mayinvolve systemic exposure or may be local administration, such as when acompound or composition is to be administered at the site of interest.Various techniques can be used for providing the subject compositions atthe site of interest, such as via injection, use of catheters, trocars,projectiles, pluronic gel, stems, sustained drug release polymers orother device which provides for internal access. Where an organ ortissue is accessible because of removal from the patient, such organ ortissue may be bathed in a medium containing the subject compositions,the subject compositions may be painted onto the organ, or may beapplied in any convenient way. The methods of the invention embraceadministration of the compounds to an organ to be donated (such as toprevent ischemia/reperfusion injury). Accordingly, organs that areremoved from one individual for transplant into another individual maybe bathed in a medium containing or otherwise exposed to a compound orcomposition as described herein.

The compounds of the invention, such as those of the formulae herein,may be administered in any suitable dosage amount, which may includedosage levels of about 0.0001 to 4.0 grams once per day (or multipledoses per day in divided doses) for adults. Thus, in certain embodimentsof this invention, a compound herein is administered at a dosage of anydosage range in which the low end of the range is any amount between 0.1mg/day and 400 mg/day and the upper end of the range is any amountbetween 1 mg/day and 4000 mg/day (e.g., 5 mg/day and 100 mg/day, 150mg/day and 500 mg/day). In other embodiments, a compound herein, isadministered at a dosage of any dosage range in which the low end of therange is any amount between 0.1 mg/kg/day and 90 mg/kg/day and the upperend of the range is any amount between 1 mg/kg/day and −32 100 mg/kg/day(e.g., 0.5 mg/kg/day and 2 mg/kg/day, 5 mg/kg/day and 20 mg/kg/day). Thedosing interval can be adjusted according to the needs of theindividual. For longer intervals of administration, extended release ordepot formulations can be used. The dosing can be commensurate withintravenous administration. For instance, the compound can beadministered, such as in a pharmaceutical composition that is amenableto intravenous administration, in an amount of between about 0.01μg/kg/min to about 100 μg/kg/min or between about 0.05 μg/kg/min toabout 95 μg/kg/min or between about 0.1 μg/kg/min to about 90 μg/kg/minor between about 1.0 μg/kg/min to about 80 μg/kg/min or between about10.0 μg/kg/min to about 70 μg/kg/min or between about 20 μg/kg/min toabout 60 μg/kg/min or between about 30 μg/kg/min to about 50 μg/kg/minor between about 0.01 μg/kg/min to about 1.0 μg/kg/min or between about0.01 μg/kg/min to about 10 μg/kg/min or between about 0.1 μg/kg/min toabout 1.0 μg/kg/min or between about 0.1 μg/kg/min to about 10 μg/kg/minor between about 1.0 μg/kg/min to about 5 μg/kg/min or between about 70μg/kg/min to about 100 μg/kg/min or between about 80 μg/kg/min to about90 μg/kg/min. In one variation, the compound is administered to anindividual, such as in a pharmaceutical composition that is amenable tointravenous administration, in an amount of at least about 0.01μg/kg/min or at least about 0.05 μg/kg/min or at least about 0.1μg/kg/min or at least about 0.15 μg/kg/min or at least about 0.25μg/kg/min or at least about 0.5 μg/kg/min or at least about 1.0μg/kg/min or at least about 1.5 μg/kg/min or at least about 5.0μg/kg/min or at least about 10.0 μg/kg/min or at least about 20.0μg/kg/min or at least about 30.0 μg/kg/min or at least about 40.0μg/kg/min or at least about 50.0 μg/kg/min or at least about 60.0μg/kg/min or at least about 70.0 μg/kg/min or at least about 80.0μg/kg/min or at least about 90.0 μg/kg/min or at least about 100.0μg/kg/min or more. In another variation, the compound is administered toan individual, such as in a pharmaceutical composition that is amenableto intravenous administration, in an amount of less than about 100.0μg/kg/min or less than about 90.0 μg/kg/min or less than about 80.0μg/kg/min or less than about 80.0 μg/kg/min or less than about 70.0μg/kg/min or less than about 60.0 μg/kg/min or less than about 50.0μg/kg/min or less than about 40.0 μg/kg/min or less than about 30.0μg/kg/min or less than about 20.0 μg/kg/min or less than about 10.0μg/kg/min or less than about 5.0 μg/kg/min or less than about 2.5μg/kg/min or less than about 1.0 μg/kg/min or less than about 0.5μg/kg/min or less than about 0.05 μg/kg/min or less than about 0.15μg/kg/min or less than about 0.1 μg/kg/min or less than about 0.05μg/kg/min or less than about 0.01 μg/kg/min.

The invention further provides kits comprising one or more compounds asdescribed herein. The kits may employ any of the compounds disclosedherein and instructions for use. The compound may be formulated in anyacceptable form. The kits may be used for any one or more of the usesdescribed herein, and, accordingly, may contain instructions for any oneor more of the stated uses (e.g., treating and/or preventing and/ordelaying the onset and/or the development of heart failure orischemia/reperfusion injury).

Kits generally comprise suitable packaging. The kits may comprise one ormore containers comprising any compound described herein. Each component(if there is more than one component) can be packaged in separatecontainers or some components can be combined in one container wherecross-reactivity and shelf life permit.

The kits may optionally include a set of instructions, generally writteninstructions, although electronic storage media (e.g., magnetic disketteor optical disk) containing instructions are also acceptable, relatingto the use of component(s) of the methods of the present invention(e.g., treating, preventing and/or delaying the onset and/or thedevelopment of heart disease or ischemia/reperfusion injury). Theinstructions included with the kit generally include information as tothe components and their administration to an individual.

The following examples are provided to illustrate various embodiments ofthe invention, and are not intended to limit the invention in anymanner.

EXAMPLES

All HPLC analysis was carried out using a CTC PAL HTS autosampler with awaters 2487 uv detector powered by an Agilent G1312A binary pump. Thefollowing method and column were used for determination of retentiontime (TR) 0-100% B [MeCN:H₂O:0.2% HCO₂H], 2.5 min gradient, 0.5 minhold, 215 nm, Atlantis dC18 2.1×50 mm, 5 μm.

All NMR were recorded on a Bruker AVANCE 400 MHz, or Bruker 500spectrometer operating at ambient probe temperature using an internaldeuterium lock. Chemical shifts are reported in parts per million (ppm)at lower frequency relative to tetramethylsilane (TMS). Standardabbreviations are used throughout (s singlet; br. s broad singlet; ddoublet; dd doublet of doublets; t triplet; q quartet; quin quintet; mmultiplet). Coupling constants are reported in Hertz (Hz).

Example 1 Preparation of Compounds of the Invention According to GeneralSynthesis of Scheme A 2-Hydroxysulfamoyl-benzoic Acid Methyl Ester(Compound 1)

A solution of potassium carbonate (3.53 g, 25.57 mmol) in water (3.6 ml)was added dropwise to a solution of hydroxylamine hydrochloride (1.76 g,25.57 mmol) in water (2.4 ml) at 0° C. maintaining an internal reactiontemperature between 5° C. and 15° C. THF (12 ml) and methanol (3 ml)were added, followed by 2-chlorosulfonyl-benzoic acid methyl ester (3 g,12.78 mmol) portionwise maintaining a temperature below 15° C. and thereaction mixture was stirred at ambient temperature until completeconsumption of the sulfonyl chloride was observed by tlc. The resultingsuspension was concentrated to remove any volatiles and the aqueoussuspension was extracted with diethyl ether (2×100 ml). The organicportion was dried over sodium sulphate, filtered and concentrated invacuo to yield the crude N-hydroxy sulfonamide. Purification wasachieved by chromatography on silica gel eluting with heptane:ethylacetate (8:2 v:v) to give the parent compound as a white solid (1.3 g,44% yield); δH (400 MHz, DMSO) 9.87 (1H, d, 3.4 Hz), 9.39 (1H, d, 3.2Hz), 7.97 (1H, m), 7.77 (2H, m), 7.68-7.72 (1H, m), 3.83 (3H, s);TR=1.43 min.

Example 2 Preparation of Compounds of the Invention According to GeneralSynthesis of Scheme A 3-Hydroxysulfamoyl-benzoic Acid (Compound 2)

A solution of potassium carbonate (0.43 g, 3.10 mmol) in water (1.2 ml)was added drop wise to a solution of hydroxylamine hydrochloride (0.21g, 3.10 mmol) in water (0.8 ml) at 0° C. maintaining an internalreaction temperature between 5° C. and 15° C. THF (4 ml) and methanol (1ml) were added, followed by 3-chlorosulfonyl-benzoic acid (0.34 g, 1.55mmol) portion wise maintaining a temperature below 15° C. and thereaction mixture was stirred at ambient temperature until completeconsumption of the sulfonyl chloride was observed by LCMS. The resultingsuspension was concentrated to remove any volatiles and the aqueoussuspension was extracted with diethyl ether (2×100 ml), and the combinedorganic layers were discarded. The aqueous layer was acidified to pH=1,adding a solution of hydrochloric acid (2N) drop wise, then extractedwith diethyl ether (2×100 ml). The organic portion was dried over sodiumsulphate, filtered and concentrated in vacuo to yield the cleanN-hydroxysulphonamide as a yellow solid (0.9 g, 27% yield); 8H (400 MHz,DMSO) 13.52 (1H, br. s.), 9.72-9.74 (2H, m), 8.38 (1H, t, 1.6 Hz), 8.23(1H, ddd, 7.9, 1.3, 1.2 Hz), 8.06 (1H, ddd, 7.8, 2.0, 1.2 Hz), 7.77 (1H,t, 7.7 Hz); TR=1.10 min.

Example 3 Preparation of Compounds of the Invention According to GeneralSynthesis of Scheme B 4-Bromo-3-(hydroxysulfamoyl)-N,N-dimethylbenzamide(Compound 6)

A solution of aqueous hydroxylamine (0.5 ml of a 50% aqueous solution,7.6 mmol) in water (1 ml) and THF (5 ml) was cooled to −5° C.2-Bromo-5-(dimethyl-4-carbonyl)benzene sulfonyl chloride was addedportionwise maintaining a temperature below about 10° C. and thereaction mixture was stirred at this temperature until completeconsumption of the sulfonyl chloride is observed by thin layerchromatography (TLC). The resulting suspension is concentrated to removeany volatiles and resulting solid was washed with diethyl ether to yieldthe title compound as a cream solid (0.36 g, 36%). δH (500 MHz, DMSO-d6)10.00 (1H, br. s), 9.88 (1H, s), 7.95 (1H, d, 1.8 Hz), 7.93 (1H, d, 8.1Hz), 7.61 (1H, dd, 8.3, 2.0 Hz), 3.00 (3H, s), 2.91 (3H, s), TR=1.19min.

Using the experimental conditions reported above and detailed in GeneralSynthesis Scheme A and B (Methods 1 and 2) and the appropriate startingmaterials, which were either commercially available or synthesised usingstandard literature conditions, the following derivatives were prepared:

Method of N-Hydroxy Compound Systematic Sulfonamide No. Name 1-H NMRT_(R) Synthesis Yield 3 4-Chloro-3- δ_(H) (400 MHz, DMSO) 13.62 1.24 A25% hydroxysulfamoyl- (1H, br. s.), 10.03 (1H, d, benzoic acid 2.9 Hz),9.88 (1H, d, 2.9 Hz), 8.50 (1H, d, 2.2 Hz), 8.16 (1H, dd, 8.3, 2.2 Hz),7.82 (1H, d, 8.3 Hz). 4 4-Bromo-3- δ_(H) (400 MHz, DMSO-d6) 1.28 A 33%hydroxysulfamoyl- 13.63 (1H, s), 10.06 (1H, s), benzoic acid 9.89 (1H,d, 1.7 Hz), 8.50 (1H, d, 1.7 Hz), 7.97-8.06 (3H, m). 5 2-Bromo-N- δ_(H)(400 MHz, DMSO-d6) 1.27 A  6% hydroxy-5- 10.02 (1H, br. s.), 9.89 (1H,(morpholine-4- s), 7.95 (1H, d, 9.3 Hz), 7.96 carbonyl)- (1H, d, 3.2Hz), 7.62 (1H, dd, benzenesulfonamide 8.2, 2.1 Hz), 3.49-3.71 (8H, m). 74-Chloro-3- δ_(H) (500 MHz, DMSO-d6) 1.18 B 79% (hydroxysulfamoyl)- 9.97(1H, s), 9.87 (1H, d, N,N- 2.1 Hz), 7.95 (1H, d, 0.8 Hz),dimethylbenzamide 7.69-7.82 (2H, m), 3.00 (3H, br. s), 2.91 (3H, br. s).8 N-Hydroxy-2- δ_(H) (400 MHz, DMSO-d6) 1.05 B 26% (morpholin-4- 10.03(1H, d, 3.4 Hz), 9.18 ylsulfonyl)benzene (1H, d, 3.4 Hz), 8.17-8.23sulfonamide (1H, m), 8.07-8.14 (1H, m), 7.90-7.97 (2H, m), 3.58- 3.64(4H, m), 3.19-3.25 (4H, m). 9 2-(Morpholin- δH (500 MHz, DMSO-d6) 0.86 B23% 4-yl)ethyl 4- 10.05 (1H, br. s.), 9.89 (1H, chloro-3- s), 8.50 (1H,s), 8.17 (1H, dd, (hydroxy- 1.2, 8.4 Hz), 7.87 (1H, d,sulfamoyl)benzoate 8.4 Hz), 4.44 (2H, t, 5.6 Hz), 3.55 (4H, t, 4.4 Hz),2.70 (2H, t, 5.7 Hz), 2.43-2.49 (4H, m). 97 4-Bromo-5- δH (500 MHz,DMSO-d6) 0.75 B 25% (hydroxysulfamoyl)- 10.05 (1H, br. s.), 9.95 (1H,N-[2- s), 8.82 (1H, t, 5.6 Hz), 7.89 (morpholin-4- (1H, s), 3.56 (4H, t,4.5 Hz), yl)ethyl]thiophene- 3.39-3.37 (2H, m), 2.46 2-carboxamide (2H,t, 6.6 Hz), 2.41 (4H, br. s.). 98 3-Bromo-N- δH (500 MHz, DMSO-d6) 1.24B 43% hydroxy-5- 10.06 (1H, br. s.), 9.97 (1H, (morpholin-4- s), 7.61(1H, s), 3.63-3.60 ylcarbonyl)thiophene- (8H, m). 2-sulfonamideSynthesis of Starting Sulfonyl Chlorides

The sulfonyl chlorides were synthesised from commercially availablestarting materials following the methods described in J. Med. Chem, 40,2017; Bioorg. Med. Chem., 2002, 639-656; Journal of Pharmacy andPharmacology, 1963, 202-211 and in Australian Journal of Chemistry,2000, 1-6. For example, certain sulfonyl chlorides can be synthesizedaccording to the following schemes:

The scheme for thiophene derivatives can be adapted to synthesizesimilar pyrrole derivatives.

2-Bromo-5-(morpholine-4-carbonyl)-benzenesulfonyl Chloride

Step 1 4-Bromo-3-chlorosulfonyl-benzoic Acid

4-Bromobenzoic acid (10 g, 49.75 mmol) was added portion wise tochlorosulfonic acid (25 ml, 373.13 mmol) at 0° C. The mixture was heatedto 130° C. for 10 hours, until complete consumption of the startingmaterial was observed by LCMS. The reaction mixture was allowed to cooldown to ambient temperature, then added drop wise to an ice/watermixture (500 ml). The precipitate was filtered and washed with coldwater (2×100 ml). The solid recovered was dissolved in diethyl ether,the solution then dried over sodium sulphate, filtered and concentratedin vacuo to afford the expected product as a beige solid (11.85 g, 79%yield); δH (400 MHz, DMSO) 8.45 (1H, d, 1.7 Hz), 7.72 (1H, d, 2.2 Hz),7.71 (1H, s); TR=2.08 min.

Step 2 4-Bromo-3-chlorosulfonyl-benzoyl Chloride

4-Bromo-3-chlorosulfonyl-benzoic acid (2 g, 6.68 mmol) was suspended intoluene (20 ml). Thionyl chloride (0.97 ml, 13.36 mmol) was added dropwise, and the mixture was heated to reflux for 14 hours under nitrogenuntil complete consumption of the carboxylic acid was observed by LCMS.The reaction mixture was concentrated under vacuum to dryness to affordthe expected acid chloride as a solid (2.12 g, 95% yield). The compoundwas used for next step without any further purification or analysis;TR=2.38.

Step 3 2-Bromo-5-(morpholine-4-carbonyl)-benzenesulfonyl Chloride

Morpholine hydrochloride (0.87 g, 7.01 mmol) was added to a solution of4-bromo-3-chlorosulfonyl-benzoyl chloride (2.12 g, 6.68 mmol) inchlorobenzene (8 ml). The reaction was heated to reflux for 2 hours,until complete consumption of the starting material was observed byLCMS. The reaction mixture was concentrated to dryness under vacuum. Theresidue was taken up in diethyl ether (20 ml), and the precipitate wasfiltered and washed with diethyl ether (2×10 ml), to afford the expectedsulfonyl chloride as a beige solid (2.34 g, 95% yield); 8H (400 MHz,DMSO) 7.90 (1H, d, 2.2 Hz), 7.64 (1H, d, 8.1 Hz), 7.25 (1H, dd, 8.1, 2.2Hz), 3.25-3.70 (8H, m); TR=2.01 min.

3-Chlorosulfonyl-4-chloro Benzoic Acid

3-Chlorosulfonyl-4-chloro benzoic acid was prepared from commerciallyavailable starting materials following Step 1 of the above detailedmethod. δ_(H) (400 MHz, DMSO-d₆) 14.03 (1H, br. s), 8.43 (1H, d, 2.2Hz), 7.84 (1H, dd, 8.1, 2.2 Hz), 7.50 (1H, d, 8.1 Hz); TR=1.81 min.

3-Chlorosulfonyl-4-bromo Benzoic Acid

3-Chlorosulfonyl-4-bromo benzoic acid was prepared from commerciallyavailable starting materials following Step 1 of the above detailedmethod. 8H (400 MHz, DMSO) 8.45 (1H, d, 1.7 Hz), 7.72 (1H, d, 2.2 Hz),7.71 (1H, s); TR=2.08 min.

2-Bromo-5-(dimethyl-4-carbonyl)benzenesulfonyl Chloride

2-Bromo-5-(dimethyl-4-carbonyl)benzenesulfonyl chloride was preparedfrom commercially available starting materials following Steps 1-3 ofthe above detailed method using dimethylamine HCl instead of morpholineHCl in Step 3. (4.35 g, 80%). δH (500 MHz, DMSO-d6) 7.89 (1H, d, 2.0Hz), 7.63 (1H, d, 8.1 Hz), 7.25 (1H, dd, 8.1, 2.2 Hz), 2.96 (3H, br. s),2.88 (3H, br. s); TR=1.78 min.

2-Chloro-5-(dimethyl-4-carbonyl)benzenesulfonyl Chloride

2-Chloro-5-(dimethyl-4-carbonyl)benzenesulfonyl chloride was preparedfrom commercially available starting materials following Steps 1-3 ofthe above detailed method using dimethylamine HCl instead of morpholineHCl in Step 3. (4.37 g, 79%). δH (500 MHz, DMSO-d6) 7.86 (1H, d, 2.0Hz), 7.43 (1H, d, 8.1 Hz), 7.34 (1H, dd, 8.1, 2.2 Hz), 2.97 (3H, br. s),2.90 (3H, br. s); TR=1.20 min.

2-(Morpholin-4-ylsulfonyl)benzene Sulfonyl Chloride

Step 1 2-(Morpholin-4-ylsulfonyl)chlorobenzene

To a solution of a 2-chlorobenzene sulfonyl chloride (10 g, 47.4 mmol)in DCM (100 ml) cooled to 0° C. was added pyridine (5.7 ml, 71.1 mmol)and morpholine (6.2 ml, 71.1 mmol) dropwise. Stirring was continued for15 minutes after which time no sulfonyl chloride was evident by LC-MS.The reaction was quenched by the addition of 2N HCl, washed with furtherportions of 2N HCl, dried over sodium sulfate, filtered and concentratedin vacuo to yield the title compound without need for additionalpurification. (11.2 g, 100%). δH (400 MHz, DMSO-d6) 7.97 (1H, dd, 7.8,1.5 Hz), 7.66-7.77 (2H, m), 7.58 (1H, ddd, 8.1, 6.7, 1.8 Hz), 3.57-3.64(4H, m), 3.11-3.18 (4H, m); TR=1.25 min.

Step 2 Benzyl-2-(morpholin-4-ylsulfonyl)benzene Sulfide

Phenyl methanethiol (7.6 ml, 43.5 mmol) was added dropwise to a 25%solution of NaOMe/MeOH (14.1 g, 65.2 mmol) in MeOH (25 ml). Theresulting thiolate was isolated by concentration in vacuo. The sodiumthiolate salt was dissolved in DMSO (65 ml), and2-(morpholin-4-ylsulfonyl)chlorobenzene (11.1 g, 43.5 mmol) addedportionwise. The reaction mixture was stirred at ambient temperature for72 h after which time no starting material was evident. The reactionmixture was poured into a 2N HCl solution (200 mL) and extracted intoDCM (3×200 ml). The organic portion was dried over sodium sulfate,filtered and concentrated in vacuo. The crude material was purified bycolumn chromatography eluting with heptane: ethyl acetate (8:2 v:v).(13.7 g, 90%), TR=1.00 min.

Step 3 2-(Morpholin-4-ylsulfonyl)benzene Sulfonyl Chloride

Chlorine gas was bubbled into a cooled (0° C.) suspension ofbenzyl-2-(morpholin-4-ylsulfonyl)benzene sulfide (7.0 g, 1.1 mmol) inconcentrated HCl (70 ml) maintaining an internal temperature below 15°C. until complete consumption of the starting material was observed(c.a. 45 minutes). After completion of the reaction, nitrogen wasbubbled into the reaction mixture for 15 minutes to remove any excesschlorine and the reaction was warmed to ambient temperature. Thereaction mixture was poured onto ice and the yellow solid formed wasfiltered and washed with ice water. The solid was trituated withheptane, filtered and washed with heptane and dried under vacuum toyield the crude sulfonyl chloride as a pale yellow solid which was useddirectly in the synthesis of Compound 8.

2-(Morpholin-4-yl)ethyl 4-chloro-3-(chlorosulfonyl)benzoate

The synthesis of 4-chloro-3-(chlorosulfonyl)benzoic acid and2-chloro-5-(dimethyl-4-carbonyl)benzenesulfonyl chloride were preparedin the same manner as detailed in the synthesis of2-bromo-5-(morpholine-4-carbonyl)-benzenesulfonyl chloride

Step 3 2-(Morpholin-4-yl)ethyl 4-chloro-3-(chlorosulfonyl)benzoate

To a solution of 4-chloro-3-(chlorosulfonyl)benzoyl chloride (1.0 g, 3.6mmol) in DCM (10 ml) was added a solution of hydroxy ethyl morpholine(443 μl, 3.6 mmol) in DCM (1 ml). The reaction mixture was stirred for12 h after which time no sulfonyl chloride was observed by LC-MS. Uponconcentration of the crude reaction mixture a solid was isolated, whichwas trituated with ether to afford the desired compound as a brownsolid. (1.54 g, 100%) IR (neat) vmax/cm⁻¹ 1724, 1174, 1450; 6H (500 MHz,DMSO-d6) 8.49 (1H, d, 1.8 Hz), 7.97 (1H, dd, 8.2, 1.9 Hz), 7.56 (1H, d,8.2 Hz), 4.38-5.00 (2H, m), 3.88-4.07 (2H, m), 3.76-3.83 (2H, m),3.56-3.62 (2H, m), 3.36-3.54 (2H, m), 3.14-3.28 (2H, m).

3-Bromo-5-{[2-(morpholin-4-yl)ethyl]carbamoyl}thiophene-2-sulfonylChloride

Step 1 4-Bromo-N-[2-(morpholin-4-yl)ethyl]thiophene-2-carboxamide

To a solution of 4-bromo-2-thiophenecarbonyl chloride (1.0 g, 4.4 mmol)in DCM (10 ml) cooled to 0° C. was added DIPEA (0.85 ml, 4.8 mml) and4-(2-aminoethyl)morpholine (0.6 ml, 4.6 mmol) dropwise. Stirring wascontinued until complete consumption of the starting material wasobserved by LC-MS. The reaction was quenched by the addition of water,which was washed with further portions of 2N HCl, dried over sodiumsulfate, filtered and concentrated in vacuo to yield the desiredcompound without need for further purification (1.6 g, 98%). δH (500MHz, CHLOROFORM-d) 7.38 (1H, d, 1.3 Hz), 7.37 (1H, d, 1.1 Hz), 3.72-3.80(4H, m), 3.53 (2H, q, 5.6 Hz), 2.59 (2H, t, 6.0 Hz), 2.48-2.54 (4H, m),TR=0.8 min.

Step 23-Bromo-5-{[2-(morpholin-4-yl)ethyl]carbamoyl}thiophene-2-sulfonylChloride

To a flask containing4-bromo-N-[2-(morpholin-4-yl)ethyl]thiophene-2-carboxamide (1.6 g, 5.1mmol) was added chlorosulfonic acid (10 ml, 30 equiv). The reaction washeated to 95° C. for 90 minutes before careful addition to ice. Theresulting solid was filtered and dried in vacuo. The crude material wastrituated with heptane:ethyl acetate to afford the title compoundwithout need for further purification (0.62 g, 29%). δH (500 MHz,DMSO-d6) 9.01 (1H, t, 5.5 Hz), 7.76 (1H, s), 3.97 (2H, d, 10.4 Hz), 3.78(2H, t, 11.5 Hz), 3.63 (2H, q, 6.0 Hz), 3.51 (2H, d, 12.3 Hz), 3.29 (2H,q, 5.7 Hz), 3.03-3.17 (2H, m), TR=1.22 min.

3-Bromo-5-(morpholin-4-ylcarbonyl)thiophene-2-sulfonyl Chloride Step 14-Bromo-N-(morpholin-4-y)thiophene-2-carboxamide

To a solution of 4-bromo-2-thiophenecarbonyl chloride (1.0 g, 4.4 mmol)in DCM (10 ml) cooled to 0° C. was added DIPEA (0.85 ml, 4.8 mml) andmorpholine (0.4 g, 4.6 mmol) dropwise. Stirring was continued untilcomplete consumption of the starting material was observed by LC-MS. Thereaction was quenched by the addition of water, which was washed withfurther portions of 2N HCl, dried over sodium sulfate, filtered andconcentrated in vacuo to yield the desired compound without need forfurther purification (1.2 g, 100%). δH (500 MHz, CHLOROFORM-d) 7.38 (1H,s), 7.18 (1H, s), 3.63-3.83 (8H, m); TR=1.52 min.

Step 2 3-Bromo-5-(morpholin-4-ylcarbonyl)thiophene-2-sulfonyl Chloride

To a flask containing 4-bromo-N-(morpholin-4-y)thiophene-2-carboxamide(1.23 g, 4.5 mmol) was added chlorosulfonic acid (9 ml, 30 equiv). Thereaction was heated to 95° C. for 90 minutes before careful addition toice. The resulting suspension was extracted into DCM (3×100 ml) and thecombined organic phases dried over sodium sulphate, filtered andconcentrated in vacuo to afford the title compound without need forfurther purification (0.54 g, 32%). δH (500 MHz, CHLOROFORM-d) 7.30 (1H,s) 3.69-3.81 (8H, m), TR=1.86 min.

Example 3 Kinetics of HNO Release

The decomposition rates of the compounds may be determined by UV-Visspectroscopy.

The decomposition of compounds may be monitored by UV-Vis spectroscopyin 0.1 M PBS buffer at pH 7.4 and 37° C. Bonner, F. T.; Ko., Y. Inorg.Chem. 1992, 31, 2514-2519.

Example 4 HNO Production Via N₂O Quantification

HNO production of the compounds was be determined by UV-Visspectroscopy.

Nitrous oxide is produced via the dimerization and dehydration of HNO,and is the most common marker for HNO production (Fukuto, J. M.;Bartberger, M. D.; Dutton, A. S.; Paolocci, N.; Wink, D. A.; Houk, K. N.Chem. Res. Toxicol. 2005, 18, 790-801). HNO, however, can also bepartially quenched by oxygen to yield a product that does not produceN₂O (See, (a) Mincione, F.; Menabuoni, L.; Briganti, F.; Mincione, G.;Scozzafava, A.; Supuran, C. T. J. Enzyme Inhibition 1998, 13, 267-284and (b) Scozzafava, A.; Supuran, C. T. J. Med. Chem. 2000, 43,3677-3687.) Using Angeli's salt (AS) as a benchmark, the relativeamounts of N₂O released from compounds are examined via GC headspaceanalysis.

The ability of compounds to donate nitroxyl at pH 7.4 in PBS buffer at37° C. was assessed. In particular, the compounds were tested and theirnitroxyl donating ability at pH 7.4 in PBS buffer at 37° C. assessed.Results are provided in Table 4 below and results for compounds 1-5 arealso illustrated in FIG. 1.

TABLE 4 Nitroxyl Donating Ability of Representative Compounds. Nitrousoxide evolved % moles N₂O/ Compound as % of Angelis salt moles of sampleAngeli's Salt 100 70.5 1 43 30.5 2 27 19 3 116 81.5 4 117 82 5 115 80.56 116 81 7 122 85 8 126 86 9 93 65 97 131 92 98 116 81

Example 5 Use of an In Vitro Model to Determine the Ability of Compoundsof the Invention to Treat, Prevent and/or Delay the Onset and/or theDevelopment of a Disease or Condition Responsive to Nitroxyl Therapy

a. Cardiovascular Diseases or Conditions

In vitro models of cardiovascular disease can also be used to determinethe ability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a cardiovasculardisease or condition in an individual. An exemplary in vitro model ofheart disease is described below.

In-vitro models could be utilized to look at vasorelaxation propertiesof the compounds. Isometric tension in isolated rat thoracic aortic ringsegment can be measured as described previously by Crawford, J. H.,Huang, J, Isbell, T. S., Shiva, S., Chacko, B. K., Schechter, A.,Darley-Usmar, V. M., Kerby, J. D., Lang, J. D., Krauss, D., Ho, C.,Gladwin, M. T., Patel, R. P., Blood 2006, 107, 566-575. Upon sacrificeaortic ring segments are excised and cleansed of fat and adheringtissue. Vessels are then cut into individual ring segments (2-3 mm inwidth) and suspended from a force-displacement transducer in a tissuebath. Ring segments are bathed at 37° C. in a bicarbonate-buffered,Krebs-Henseleit (K-H) solution of the following composition (mM): NaCl118; KCl 4.6; NaHCO₃ 27.2; KH₂PO₄ 1.2; MgSO₄ 1.2; CaCl₂ 1.75; Na₂EDTA0.03; and glucose 11.1 and perfused continuously with 21% O₂/5% CO₂/74%N₂. A passive load of 2 g is applied to all ring segments and maintainedat this level throughout the experiments. At the beginning of eachexperiment, indomethacin-treated ring segments are depolarized with KCl(70 mM) to determine the maximal contractile capacity of the vessel.Rings are then washed extensively and allowed to equilibrate. Forsubsequent experiments, vessels are submaximally contracted (50% of KClresponse) with phenylephrine (PE, 3×10⁻⁸-10⁻⁷ M), and L-NMMA, 0.1 mM, isalso added to inhibit eNOS and endogenous NO production. After tensiondevelopment reaches a plateau, nitroxyl donating compounds are addedcumulatively to the vessel bath and effects on tension monitored.

In vitro models can be utilized to determine the effects of nitroxyldonating compounds in changes in developed force and intracellularcalcium in heart muscles. Developed force and intracellular calcium canbe measured in rat trabeculae from normal or diseased (i.e. rats withcongestive heart failure or hypertrophy) as described previously (Gao WD, Atar D, Backx P H, Marbán E. Circ Res. 1995; 76:1036-1048). Rats(Sprague-Dawley, 250-300 g) are used in these experiments. The rats areanesthetized with pentobarbital (100 mg/kg) via intra-abdominalinjection, the heart exposed by mid-sternotomy, rapidly excised andplaced in a dissection dish. The aorta is cannulated and the heartperfused retrograde (˜15 mM/min) with dissecting Krebs-Henseleit (H-K)solution equilibrated with 95% O₂ and 5% CO₂. The dissecting K-Hsolution is composed of (mM): NaCl 120, NaHCO₃ 20, KCl 5, MgCl 1.2,glucose 10, CaCl₂ 0.5, and 2,3-butanedione monoximine (BDM) 20, pH7.35-7.45 at room temperature (21-22° C.). Trabeculae from the rightventricle of the heart are dissected and mounted between a forcetransducer and a motor arm and superfused with normal K-H solution (KCl,5 mM) at a rate of ˜10 ml/min and stimulated at 0.5 Hz. Dimensions ofthe muscles are measured with a calibration reticule in the ocular ofthe dissection microscope (×40, resolution ˜10 μm).

Force is measured using a force transducer system and is expressed inmilli newtons per square millimeter of cross-sectional area. Sarcomerelength is measured by laser diffraction. Resting sarcomere length is setat 2.20-2.30 μm throughout the experiments.

Intracellular calcium is measured using the free acid form of fura-2 asdescribed in previous studies (Gao et al., 1994; Backx et al., 1995; Gaoet al., 1998). Fura-2 potassium salt is microinjected iontophoreticallyinto one cell and allowed to spread throughout the whole muscle (via gapjunctions). The tip of the electrode (˜0.2 μm in diameter) is filledwith fura-2 salt (1 mM) and the remainder of the electrode was filledwith 150 mM KCl. After a successful impalement into a superficial cellin non-stimulated muscle, a hyperpolarizing current of 5-10 nA is passedcontinuously for ˜15 min. Fura-2 epifluorescence is measured by excitingat 380 and 340 nm. Fluorescent light is collected at 510 nm by aphotomultiplier tube. The output of photomultiplier is collected anddigitized. Ryanodine (1.0 μM) is used to enable steady-state activation.After 15 min of exposure to ryanodine, different levels of tetanizationsare induced briefly (˜4-8 seconds) by stimulating the muscles at 10 Hzat varied extracellular calcium (0.5-20 mM). All experiments areperformed at room temperature (20-22° C.).

b. Diseases or Conditions Implicating Ischemia/Reperfusion.

In vitro models can also be used to determine the ability of any of thecompounds described herein to treat, prevent and/or delay the onsetand/or the development of a disease or condition implicatingischemia/reperfusion injury in an individual.

Example 6 Use of In Vivo and/or Ex Vivo Models to Determine the Abilityof Compounds of the Invention to Treat, Prevent and/or Delay the Onsetand/or the Development of a Disease or Condition Responsive to NitroxylTherapy

a. Cardiovascular Diseases or Conditions.

In vivo models of cardiovascular disease can also be used to determinethe ability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a cardiovasculardisease or condition in an individual. An exemplary animal model ofheart disease is described below.

In vivo cardiovascular effects obtained with a nitroxyl donor compoundmay be assessed in a control (normal) dog. The study is conducted inadult (25 kg) mongrel (male) dogs chronically instrumented for conscioushemodynamic analysis and blood sampling, as previously described(Katori, T.; Hoover, D. B.; Ardell, J. L.; Helm, R. H.; Belardi, D. F.;Tocchetti, C. G.; Forfia, P. R.; Kass, D. A.; Paolocci, N. Circ. Res.96(2): 2004). Micromanometer transducers in the left ventricle providepressure, while right atrial and descending aortic catheters providefluid-pressures and sampling conduits. Endocardial sonomicrometers(anteriorposterior, septal-lateral) measure short-axis dimensions, apneumatic occluder around the inferior vena cave facilitated pre-loadmanipulations for pressure-relation analysis. Epicardial pacing leadsare placed on the right atrium, and another pair is placed on the rightventricle free wall linked to a permanent pacemaker to induce rapidpacing-cardiac failure. After 10 days of recovery, animals are evaluatedat baseline sinus rhythm and with atrial pacing (120-160 bpm).Measurements include conscious hemodynamic recordings for cardiacmechanics.

Compounds of the invention are administrated to a healthy control dog atthe dose of 1-5 μg/kg/min and the resulting cardiovascular data isobtained.

Demonstration that a compound of the invention improves cardiachemodynamics in hearts with congestive failure: After completingprotocols under baseline conditions, congestive heart failure is inducedby tachypacing (210 bpm×3 weeks, 240 bpm×I week), as previouslydescribed (Katori, T.; Hoover, D. B.; Ardell, J. L.; Helm, R. H.;Belardi, —37 D. F.; Tocchetti, C. G.; Forfia, P. R.; Kass, D. A.;Paolocci, N. Circ. Res. 96(2): 2004). Briefly, end-diastolic pressureand +dP/dt,max are measured weekly to monitor failure progression. Whenanimals demonstrate a rise in EDP more than 2×, and dp/dt,max of >50%baseline, they are deemed ready for congestive heart failure studies.

The values for test compounds are obtained after 15 min continuous i.v.infusion (2.5 or 1.25 μg/kg/min) in control and heart failurepreparations, respectively, both in the absence and in the presence ofvolume restoration. For comparison, the same hemodynamic measurementsare obtained with AS in heart failure preparations.

b. Diseases or Conditions Implicating Ischemia/Reperfusion.

Ex-vivo models of ischemia/reperfusion can also be used to determine theability of any of the compounds described herein to treat, preventand/or delay the onset and/or the development of a disease or conditionimplicating ischemia/reperfusion injury in an individual. An exemplaryex vivo model of ischemia/reperfusion injury is described below.

Male Wistar rats are housed in identical cages and allowed access to tapwater and a standard rodent diet ad libitum. Each animal is anesthetizedwith 1 g/kg urethane i.p. 10 min after heparin (2,500 U, i.m.)treatment. The chest is opened, and the heart is rapidly excised, placedin ice-cold buffer solution and weighed. Isolated rat hearts areattached to a perfusion apparatus and retrogradely perfused withoxygenated buffer solution at 37° C. The hearts are instrumented aspreviously described in Rastaldo et al., “P-450 metabolite ofarachidonic acid mediates bradykinin-induced negative inotropic effect,”Am. J. Physiol., 280:H2823-H2832 (2001), and Paolocci et al.“cGMP-independent inotropic effects of nitric oxide and peroxynitritedonors: potential role for nitrosylation,” Am. J. Physiol., 279:H1982-H1988 (2000). The flow is maintained constant (approximately 9mL/min/g wet weight) to reach a typical coronary perfusion pressure of85-90 mm Hg. A constant proportion of 10% of the flow rate is applied bymeans of one of two perfusion pumps (Terumo, Tokyo, Japan) using a 50 mLsyringe connected to the aortic cannula. Drug applications are performedby switching from the syringe containing buffer alone to the syringe ofthe other pump containing the drug (nitroxyl donating compound)dissolved in a vehicle at a concentration 10× to the desired finalconcentration in the heart. A small hole in the left ventricular wallallows drainage of the thebesian flow, and a polyvinyl-chloride balloonis placed into the left ventricle and connected to an electromanometerfor recording of left ventricular pressure (LVP). The hearts areelectrically paced at 280-300 bpm and kept in a temperature-controlledchamber (37° C.). Coronary perfusion pressure (CPP) and coronary floware monitored with a second electromanometer and an electromagneticflow-probe, respectively, both placed along the perfusion line. Leftventricular pressure, coronary flow and coronary perfusion pressure arerecorded using a TEAC R-71 recorder, digitized at 1000 Hz and analyzedoff-line with DataQ-Instruments/CODAS software, which allowquantification of the maximum rate of increase of LVP during systole(dP/dt_(max)).

Hearts are perfused with Krebs-Henseleit solution gassed with 95% O₂ and5% CO₂ of the following composition: 17.7 mM sodium bicarbonate, 127 mMNaCl, 5.1 mM KCl, 1.5 mM CaCl₂, 1.26 mM MgCl₂, 11 mM D-glucose,supplemented with 5 μg/mL lidocaine.

Experimental Compounds. The nitroxyl donors are diluted in bufferimmediately prior to use.

Experimental Protocols. Hearts are allowed to stabilize for 30 min, andbaseline parameters are recorded. Typically, coronary flow is adjustedwithin the first 10 min and kept constant from thereon. After 30 minstabilization, hearts are randomly assigned to one of the treatmentgroups, and subjected to 30 min global, no-flow ischemia, followed by 30min of reperfusion (I/R). Pacing of the hearts is stopped at thebeginning of the ischemic period and restarted after the third minute ofreperfusion.

Hearts in a control group are perfused with buffer for an additional 29min after stabilization. Treated hearts are exposed to a nitroxyl donor(e.g., 1 μM final concentration for about 20 min followed by a 10 minbuffer wash-out period).

In all hearts pacing is suspended at the onset of ischemia and restarted3 minutes following reperfusion. As isolated heart preparations maydeteriorate over time (typically after 2-2.5 hrs perfusion), the re-flowduration is limited to 30 min in order to minimize the effects producedby crystalloid perfusion on heart performance, and consistently withother reports.

Assessment of ventricular function. To obtain the maximal developed LVP,the volume of the intra-ventricular balloon is adjusted to anend-diastolic LVP of 10 mm Hg during the stabilization period, asreported in Paolocci, supra, and Hare et al., “Pertussis toxin-sensitiveG proteins influence nitric oxide synthase III activity and proteinlevels in rat hearts,” J. Clin. Invest., 101: 1424-31 (1998). Changes indeveloped LVP, dP/dt_(max) and the end-diastolic value induced by theI/R protocol are continuously monitored. The difference between theend-diastolic LVP (EDLVP) before the end of the ischemic period andduring pre-ischemic conditions is used as an index of the extent ofcontracture development. Maximal recovery of developed LVP anddP/dt_(max) during reperfusion is compared with respective pre-ischemicvalues.

Assessment of myocardial injury. Enzyme release is a measure of severemyocardial injury that has yet to progress to irreversible cell injury.Samples of coronary effluent (2 mL) are withdrawn with a catheterinserted into the right ventricle via the pulmonary artery. Samples aretaken immediately before ischemia and at 3, 6, 10, 20 and 30 min ofreperfusion. LDH release is measured as previously described byBergmeyer & Bemt, “Methods of Enzymatic Analysis,” Verlag Chemie (1974).Data are expressed as cumulative values for the entire reflow period.

To corroborate the data relative to myocardial injury, determined by LDHrelease, infarct areas are also assessed in a blinded fashion. At theend of the course (30 min reperfusion), each heart is rapidly removedfrom the perfusion apparatus, and the LV dissected into 2-3 mmcircumferential slices. Following 15 min of incubation at 37° C. in 0.1%solution of nitro blue tetrazolium in phosphate buffer as described inMa et al., “Opposite effects of nitric oxide and nitroxyl onpostischemic myocardial injury,” Proc. Natl. Acad. Sci., 96:14617-14622(1999), unstained necrotic tissue is separated from the stained viabletissue. The areas of viable and necrotic tissue are carefully separateby and independent observer who is not aware of the origin of thehearts. The weight of the necrotic and non-necrotic tissues is thendetermined and the necrotic mass expressed as a percentage of total leftventricular mass.

Data may be subjected to statistical methods such as ANOVA followed bythe Bonferroni correction for post hoc t tests.

Example 7 Use of Human Clinical Trials to Determine the Ability toCombination Therapies of the Invention to Treat, Prevent and/or Delaythe Onset and/or the Development of a Disease or Condition Responsive toNitroxyl Therapy

If desired, any of the compounds described herein can also be tested inhumans to determine the ability of the compound to treat, prevent and/ordelay the onset and/or the development of a disease or conditionresponsive to nitroxyl therapy. Standard methods can be used for theseclinical trials. In one exemplary method, subjects with such a diseaseor condition, such as congestive heart failure, are enrolled in atolerability, pharmacokinetics and pharmacodynamics phase I study of atherapy using the compounds of the invention in standard protocols. Thena phase II, double-blind randomized controlled trial is performed todetermine the efficacy of the compounds using standard protocols.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it is apparent to those skilled in the art that certainminor changes and modifications will be practiced. Therefore, thedescription and examples should not be construed as limiting the scopeof the invention.

All references, publications, patents, and patent applications disclosedherein are hereby incorporated by reference in their entireties.

1. A compound of formula (I)

wherein: R¹ is H; R² is H; R³, R⁴, R⁵, R⁶ and R⁷ are independentlyselected from the group consisting of H, halo, alkylsulfonyl,N-hydroxylsulfonamidyl, perhaloalkyl, nitro, aryl, cyano, alkoxy,perhaloalkoxy, alkyl, substituted aryloxy, alkylsulfanyl, alkylsulfinyl,heterocycloalkyl, substituted heterocycloalkyl, dialkylamino,cycloalkoxy, cycloalkylsulfanyl, arylsulfanyl, arylsulfinyl, carboxyl,carboxyl ester, acylamino and sulfonylamino, provided that at least oneof R³, R⁴, R⁵, R⁶ and R⁷ is carboxyl, carboxyl ester, acylamino orsulfonylamino; and provided that when one of R³, R⁴, R⁵, R⁶ and R⁷ iscarboxyl, —C(O)O-alkyl or SO₂NH₂, then at least one of the remaining R³,R⁴, R⁵, R⁶ and R⁷ is a moiety other than H or halo; or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1,wherein R² is H.
 3. The compound of claim 1, wherein at least one of R³and R⁷ is other than H.
 4. The compound of claim 1, wherein at least oneof R³ and R⁷ is an electron withdrawing group.
 5. The compound of claim1, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ is carboxyl,—COO-alkyl, —C(O)NH₂, —C(O)NR_(a)R_(b) where R_(a) is hydrogen and R_(b)is alkyl, —C(O)NR_(a)R_(b) where R_(a) and R_(b) are independentlyalkyl, —C(O)NR_(a)R_(b) where R_(a) and R_(b) are taken together withthe nitrogen to which they are attached to form a heterocyclic orsubstituted heterocyclic ring, —SO₂NH₂, —SO₂NR-alkyl where R ishydrogen, —SO₂NR-alkyl where R is alkyl, or —SO₂NR₂ where the two Rgroups are taken together with the nitrogen to which they are attachedto form a heterocyclic or substituted heterocyclic ring.
 6. Apharmaceutical composition comprising a compound of claim 1 and apharmaceutically acceptable carrier.
 7. A kit comprising a compound ofclaim 1 and instructions for use in the treatment of a disease orcondition that is responsive to nitroxyl therapy.
 8. The compound ofclaim 1, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ is carboxylester, acylamino or sulfonylamino.
 9. The compound of claim 1, whereinat least one of R³, R⁴, R⁵, R⁶ and R⁷ is —COO-alkyl.
 10. The compound ofclaim 1, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ is —S(O)₂alkyl.11. The compound of claim 1, wherein at least one of R³, R⁴, R⁵, R⁶ andR⁷ is —C(O)NR_(a)R_(b) where R_(a) is hydrogen and R_(b) is alkyl,—C(O)NR_(a)R_(b) where R_(a) and R_(b) are independently alkyl,—C(O)NR_(a)R_(b) where R_(a) and R_(b) are taken together with thenitrogen to which they are attached to form a heterocyclic orsubstituted heterocyclic ring, —SO₂NH₂, —SO₂NR-alkyl where R ishydrogen, —SO₂NR-alkyl where R is alkyl, or —SO₂NR₂ where the two Rgroups are taken together with the nitrogen to which they are attachedto form a heterocyclic or substituted heterocyclic ring.
 12. Thecompound of claim 1, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ is—C(O)NR_(a)R_(b) where R_(a) and R_(b) are independently alkyl.
 13. Thecompound of claim 1, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ iscarboxyl.
 14. The compound of claim 1, wherein the compound is selectedfrom compounds 1 to 72 in Table
 1. 15. The pharmaceutical composition ofclaim 6, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ in the compoundof formula I is carboxyl ester, acylamino or sulfonylamino.
 16. Thepharmaceutical composition of claim 6, wherein at least one of R³, R⁴,R⁵, R⁶ and R⁷ in the compound of formula I is —COO-alkyl.
 17. Thepharmaceutical composition of claim 6, wherein at least one of R³, R⁴,R⁵, R⁶ and R⁷ in the compound of formula I is —S(O)₂alkyl.
 18. Thepharmaceutical composition of claim 6, wherein at least one of R³, R⁴,R⁵, R⁶ and R⁷ in the compound of formula I is —C(O)NR_(a)R_(b) whereR_(a) is hydrogen and R_(b) is alkyl, —C(O)NR_(a)R_(b) where R_(a) andR_(b) are independently alkyl, —C(O)NR_(a)R_(b) where R_(a) and R_(b)are taken together with the nitrogen to which they are attached to forma heterocyclic or substituted heterocyclic ring, —SO₂NH₂, —SO₂NR-alkylwhere R is hydrogen, —SO₂NR-alkyl where R is alkyl, or —SO₂NR₂ where thetwo R groups are taken together with the nitrogen to which they areattached to form a heterocyclic or substituted heterocyclic ring. 19.The pharmaceutical composition of claim 6, wherein at least one of R³,R⁴, R⁵, R⁶ and R⁷ in the compound of formula I is —C(O)NR_(a)R_(b) whereR_(a) and R_(b) are independently alkyl.
 20. The pharmaceuticalcomposition of claim 6, wherein at least one of R³, R⁴, R⁵, R⁶ and R⁷ inthe compound of formula I is carboxyl.
 21. The pharmaceuticalcomposition of claim 6, wherein the compound of formula I is selectedfrom compounds 1 to 72 in Table 1.